Instrument-Based Transfection Methods

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en-us LUSONV30E Instrument based Instrument-Based Transfection Methods /webroot/web/html/lsr/solutions/technologies/transfection <p>Transfection of cells can be accomplished by various methods, including chemical, biological, and instrument-based. This section provides an overview of the different instrument-based transfection methods available, discusses how they work, and describes their pros and cons.</p> <p><strong>Related Topics</strong>: <a href="/evportal/destination/solutions?catID=LUSOOP49">Chemical- and Viral-Based Transfection Methods</a>, <a href="/evportal/destination/solutions?catID=LUSOPS84">Posttransfection Analysis of Cells</a> and <a href="/evportal/destination/solutions?catID=LUSOLB470">Cell Counting Methods</a>.</p> Overview of Instrument-Based Transfection Methods <p>Transfection can be accomplished using chemical, biological, or physical methods. Common methods include electroporation, the use of virus vectors, lipofection, and biolistics. Many types of genetic material, including plasmid DNA, siRNA, proteins, dyes, and antibodies may be transfected using any of these methods. However, no single method can be applied to all types of cells; transfection efficiencies and cytotoxicity may vary dramatically and depend on the method, cell type being utilized, and types of experiments being performed. Therefore, to obtain high efficiencies, all relevant factors should be considered in planning and selecting the appropriate transfection method.</p> <table class="pd_table pd_gridlines txttop" border="0"> <tbody> <tr class="pd_colorbackground"> <td><strong>Method</strong></td> <td><strong>Function</strong></td> <td><strong>Pros</strong></td> <td><strong>Cons</strong></td> <td><strong>Cells</strong></td> <td><strong>Products</strong></td> </tr> <tr> <td valign="top">Electroporation</td> <td valign="top">Nucleic acids or other molecules are introduced into cells by creating transient pores in the plasma membrane using an electric pulse</td> <td valign="top">Nonchemical method that doesn't seem to alter the biological structure or function of the target cells<br /><br /> Easy to perform<br /><br /> High efficiency<br /><br /> Can be applied to a wide range of cell types</td> <td valign="top">Cell mortality (if using suboptimal conditions)</td> <td valign="top">Eukaryotic cells (primary, stem cells)<br /><br /> Prokaryotic cells (bacteria, yeast)<br /><br />Plant protoplasts</td> <td valign="top"><a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=b1a35eb3-d55c-47b3-aaf3-95e4d1d85848">Gene Pulser Xcell<sup>&trade;</sup> electroporation system</a><br /> <br /> <a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=fae0f825-da45-4b02-aa33-73781dda6171">Gene Pulser MXcell<sup>&trade;</sup> electroporation system</a><br /> <br /> <a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=83527990-34fb-4b33-b955-ca53b57bf8b9">MicroPulser<sup>&trade;</sup> electroporator</a></td> </tr> <tr> <td valign="top">Biolistic particle delivery</td> <td valign="top">Delivery of nucleic acids into cells via high-velocity nucleic acid-coated microparticles</td> <td valign="top">Simple, rapid, versatile technique<br /><br /> Targeted intracellular gene delivery<br /><br /> Cell type independent<br /><br /> Uses small amounts of DNA<br /><br /> Delivers single or multiple genes<br /><br /> No carrier DNA needed<br /><br /> Can deliver large DNA fragments<br /><br /> No extraneous genes or proteins delivered<br /><br /> Requires little manipulation of cells<br /><br /> High reproducibility</td> <td valign="top">Generally lower efficiency compared to electroporation or viral or lipid mediated transfection<br /><br /> Limited bacterial transfection data<br /><br /> Requires the preparation of microparticles<br /><br /> Instrument cost<br /><br /> Requires purchase agreement</td> <td valign="top">Plant<br /><br /> Primary cells<br /><br /> Tissue<br /><br /> In vivo applications</td> <td valign="top"><a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=42e9d6be-369a-49f8-8fbb-281a0fea6df8">Helios<sup>&reg;</sup> gene gun system</a><br /> <br /> <a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=1730e08d-f43a-46ea-b7f3-7b35c04c36eb">PDS-1000/He and Hepta<sup>&trade;</sup> systems</a></td> </tr> <tr> <td valign="top">Microinjection</td> <td valign="top">Direct injection of naked DNA</td> <td valign="top">Can be used for many animals</td> <td valign="top">Laborious (one cell at a time)<br /><br /> Technically demanding and costly</td> <td valign="top">Eukaryotic cells</td> <td valign="top">&nbsp;</td> </tr> <tr> <td valign="top">Laserfection/<br /> optoinjection</td> <td valign="top">Uses laser light to transiently permeabilize a large number of cells in a very short time</td> <td valign="top">Very efficient<br /><br /> Works with many cell types<br /><br /> Few cell manipulations needed<br /><br /></td> <td valign="top">Requires cell to be attached<br /><br /> Expensive laser equipment required</td> <td valign="top">Attached cells</td> <td valign="top">&nbsp;</td> </tr> </tbody> </table> <div class="top"><a href="#helptop">Back to Top</a></div> Electroporation <p><img src="/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img1.jpg" alt="" width="245" height="624" align="left" /></p> <ul style="list-style-type: none;"> <li style="margin-bottom: 125px;">1. Electroporation exposes a cell to a high-intensity electric field that temporarily destabilizes the membrane</li> <li style="margin-bottom: 135px;">2. During this time the membrane is highly permeable to exogenous molecules present in the surrounding media</li> <li style="margin-bottom: 145px;">3. DNA then moves into the cell through these holes</li> <li>4. When the field is turned off, the pores in the membrane reseal, enclosing the DNA inside</li> </ul> <p class="caption" style="clear: both;"><strong>Electroporation of cells.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> Biolistic Particle Delivery <p>Biolistics is the delivery of nucleic acids into cells by firing nucleic acid-coated microparticles into them.</p> <p><strong>Helios Gene Gun</strong></p> <ul> <li>For in situ, in vivo and in vitro transformations</li> <li>Applications for animals, plants, cell culture, nematodes, yeast and bacteria</li> <li>Pressure range 100&ndash;600 psi enables fine-tuning of penetration</li> <li>Highly portable can be used in the field</li> <li>Small target area for accurate targeting</li> </ul> <p><img src="/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img2.jpg" alt="" width="560" height="270" /></p> <p class="caption"><strong>Biolistic particle delivery workflow.</strong></p> <p><strong>PDS-1000/He Biolistic Particle Delivery System</strong></p> <ul> <li>For in vitro, ex vivo (and in vivo for some plants and microbes)</li> <li>Applications for animal cell and organ culture, plant cell culture and explants, pollen, insects, algae, fungi and bacteria</li> <li>Pressure range 450&ndash;2200 psi gives flexibility and penetration &mdash; ideal for plant applications</li> <li>Large target area &mdash; more cells can be transformed</li> </ul> <p><strong>Procedure</strong></p> <ol> <li>DNA-coated microcarriers (thin plastic disk) are spread over the central area of that disk using a pipette tip.</li> <li>Disk loaded with the DNA-coated particles is placed into a holder inside the PDS-1000 system.</li> <li>The system uses high pressure helium, released by a rupture disk, and a partial vacuum, to propel the macrocarrier sheet loaded with DNA-coated gold macrocarriers toward the target cells.</li> <li>Macrocarrier is halted after a short distance by a stopping screen.</li> <li>DNA-coated particles continue traveling toward the target to penetrate the cells.</li> <li>Sample chamber is subjected to a partial vacuum, from 15 to 29 in. of mercury, depending on the target cells.</li> </ol> <p>&nbsp;</p> <p><img src="/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img3.jpg" alt="" width="560" height="224" /></p> <p class="caption"><strong>Workflow for delivery using PDS-1000/He system.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> Microinjection <ul> <li>Direct injection of naked DNA</li> <li>Laborious (one cell at a time)</li> <li>Technically demanding and costly</li> <li>Can be used for many animals</li> </ul> <p><img src="http://qaevn.bio-rad.com/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img4.jpg" alt="" width="532" height="172" /></p> <p class="caption"><strong>Microinjection of particles.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> Laserfection/Optoinjection <ul> <li>This procedure uses laser light to transiently permeabilize a large number of cells in a very short time</li> <li>Various substances, including ions, small molecules, dextrans, short interfering RNAs (siRNAs), plasmids, proteins, and semiconductor nanocrystals can be efficiently optoinjected into numerous cell types</li> </ul> <p><img src="http://qaevn.bio-rad.com/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img5.jpg" alt="" width="532" height="144" /></p> <p class="caption"><strong>Workflow for laserfection.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> Transfection Protocol Library <p> <script type="text/javascript"><!-- // popupwin function popIt(url,w,h,r){ var mydate = new Date(); wname=''+mydate.getMonth()+mydate.getDate()+mydate.getHours()+mydate.getMinutes()+mydate.getSeconds(); if (w && h && r) { popwin = window.open(url,"popwin"+wname,"height="+h+",width="+w+",status=1,scrollbars=1,location=1,menubar=1,resizable"); } else { popwin = window.open(url,"popwin"+wname,"height="+h+",width="+w+",status=1,scrollbars=1,location=0,resizable"); } } // --></script> </p> <p>The transfection protocol online library contains protocols obtained from the literature, developed by Bio-Rad scientists, or submitted by scientists like you. <a onclick="popIt('/genetransferprotocols',1300,700);return false" href="#" target="_blank">Browse protocols</a> to view our library and find your starting point or submit a protocol by clicking the proper technology.</p> <div class="top"><a href="#helptop">Back to Top</a></div> Further Reading <p>Belyansteva IA (2009). Helios Gene Gun-mediated transfection of the inner ear sensory epithelium. Methods Mol Biol 493, 103&ndash;123. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/18839344" target="_blank">18839344</a></p> <p>Benediktsson AM et al. (2005). Ballistic labeling and dynamic imaging of astrocytes in organotypic hippocampal slice cultures. J Neurosci Methods 141, 41&ndash;53. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/15585287" target="_blank">15585287</a></p> <p>Eizema K et al. (2000). Endothelin-1 responsiveness of a 1.4 kb phospholamban promoter fragment in rat cardiomyocytes transfected by the gene gun. J Mol Cell Cardiol 32, 311&ndash;321. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/10722806" target="_blank">10722806</a></p> <p>Fujiki R et al. (2009). GlcNAcylation of a histone methyltransferase in retinoic-acid-induced granulopoiesis. Nature 459, 455&ndash;459. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/19377461" target="_blank">19377461</a></p> <p>Gildea JJ et al. (2009). Caveolin-1 and dopamine-mediated internalization of NaKATPase in human renal proximal tubule cells. Hypertension 54, 1070&ndash;1076. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/19752292" target="_blank">19752292</a></p> <p>Helledie T et al. (2008). A simple and reliable electroporation method for human bone marrow mesenchymal stem cells. Stem Cells Dev 17, 837&ndash;848. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/18752428" target="_blank">18752428</a></p> <p>Hockemeyer D et al. (2009). Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases. Nat Biotechnol 27, 851&ndash;857. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/19680244" target="_blank">19680244</a></p> <p>Huang B et al. (2008). RNA interference-mediated in vivo silencing of fas ligand as a strategy for the enhancement of DNA vaccine potency. Hum Gene Ther 19, 763&ndash;773. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/18627219" target="_blank">18627219</a></p> <p>Shimamura K et al. (2007). Generation of secondary small interfering RNA in cell-autonomous and non-cell autonomous RNA silencing in tobacco. Plant Mol Biol 63, 803&ndash;813. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/17225952" target="_blank">17225952</a></p> <p>Su L et al. (2009). Neural stem cell differentiation is mediated by integrin beta4 in vitro. Int J Biochem Cell Biol 41, 916&ndash;924. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/18834954" target="_blank">18834954</a></p> <p>Tseng CN et al. (2013). A method to identify RNA A-to-I editing targets using I-specific cleavage and exon array analysis. Mol Cell Probes 7, 38&ndash;45. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/22960667" target="_blank">22960667</a></p> <p>Wirth MJ and Wahle P (2003). Biolistic transfection of organotypic cultures of rat visual cortex using a handheld device. J Neurosci Methods 125, 45&ndash;54. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/12763229" target="_blank">12763229</a></p> <p>Zhang G and Selzer ME (2001). In vivo transfection of lamprey brain neurons by gene gun delivery of DNA. Exp Neurol 167, 304&ndash;311. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/11161618" target="_blank">11161618</a></p> <div class="top"><a href="#helptop">Back to Top</a></div> Selection guides <table id="carttablealigned" class="literature_table" style="height: auto; width: 583px;" border="0" cellspacing="0" cellpadding="0"> <tbody> <tr> <td width="100">6176</td> <td width="350">Electroporation Systems Overview</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6176.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6177</td> <td width="350">Biolistic Particle Delivery Systems</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6177.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6178</td> <td width="350">Recommended Biolistic System by Cell Types</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6178.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6179</td> <td width="350">Lipid Transfection Reagents Selection Guide</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6179.pdf" target="_blank"><span>Click to download</span></a></td> </tr> </tbody> </table> <div class="videowrap vwrap_last"> <div class="videoImg"><a title="Gene Pulser Xcell Electroporation System: Components, Application, and Troubleshooting" onclick="window.open('/webroot/web/movies/lsr/support/Gene_Pulser_Xcell.htm', 'StatusBar', 'toolbar=no, resizable=no, scrollbars=no, width=810, height=575');" href="javascript:void(0);"><img style="border:none;" src="https://www.bio-rad.com/webroot/web/images/lsr/support/tutorials/global/xcell_tn.png" alt="" /></a></div> <div class="videoDesc"><a title="Gene Pulser Xcell Electroporation System: Components, Application, and Troubleshooting" onclick="window.open('/webroot/web/movies/lsr/support/Gene_Pulser_Xcell.htm', 'StatusBar', 'toolbar=no, resizable=no, scrollbars=no, width=810, height=575');" href="javascript:void(0);">Gene Pulser Xcell&trade; Electroporation System: Components, Application, and Troubleshooting</a><br /> This tutorial highlights the main components and features of the Gene Pulser Xcell system. It provides information about system installation and the setup of electroporation experiments, including important troubleshooting tips and answers to frequently asked questions. Ordering information for system components and accessories is also provided.</div> <div class="clear">&nbsp;</div> </div> 5448 MicroPulser Electroporator Flier, Rev A 5448 /webroot/web/pdf/lsr/literature/Bulletin_5448.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif No MicroPulser Electroporator Flier, Rev A 5448 165-2082, 165-2091, 1652081, 1652083, 1652092, ark, cuvettes, escherichia coli, fungi, schizosaccharomyces pombe, staphylococcus aureus, 165-2092, cuvette, e, electroporate, genepulser, microorganism, 165-2083, 1652082, 1652093, a, agrobacterium tumefaciens, d, microorganisms, transform, yeast, charge, electroporation, electropotators, LIT5448, micro pulser, pichia pastons, saccharomyces ceneisiae, transfection, transformation, 165-2089, 1652086, 1652088, chamber, p, s, 1652091, arc-quenching, arq, transfect, 165-2081, 165-2086, 165-2093, 1652089, bacteria, dictyostelium discoideum, micropulse, pulses, 165-2088, are quenching, field strength, gene transfer, programmable, pulse 5542 5542 Electroporation Systems Brochure, Rev A H /webroot/web/pdf/lsr/literature/Bulletin_5542.pdf Literature PDF Brochures and Specifications /webroot/web/images/general/icons/icon_pdf.gif No Electroporation Systems Brochure, Rev A Life Science 5542 165-2092, 165-2660, 165-2662, cell, cuvette, gene-pulser, genepulser, mammalian, shockpod, 165-2666, 1652091, 1652661, acids, arc-quenching, arq, electroporating, eukaryotic, fungal, micropulser, prokaryotic, transfect, 165-2082, 165-2091, 1652081, 1652083, 1652092, 1652100, 1652667, 1652669, arc quenching, ark, cuvet, cuvettes, Escherichia coli, eucaryotic, fungi, micro-pulse, shock-pod, 165-2081, 165-2086, 165-2093, 165-2661, 1652089, bacteria, cells, e., e.coli, electroporator, micropulse, pulsetrac, yeasts, 165-2083, 165-2100, 1652082, 1652093, 1652668, bacterial, dna, Gene-pulser xcell, pulse trac, rna, transform, yeast, 165-2089, 165-2667, 165-2669, 1652086, 1652088, 1652666, electroporators, fungus, saccharomyces cerevisiae, transfirmation, 165-2088, 165-2668, electroporaters, eucaryotes, eukaryotes, excel, gene transfer, prokaryotes, 1652660, 1652662, cuvets, electroporater, LIT5542, micro pulser, nucleic acid, track, transfection, transformation 5582 Gene Pulser Electroporation Buffer Product Information Sheet, Rev A 5582 /webroot/web/pdf/lsr/literature/Bulletin_5582.pdf Literature PDF Product_Information_Sheets /webroot/web/images/general/icons/icon_pdf.gif No Gene Pulser Electroporation Buffer Product Information Sheet, Rev A 5582 difficult-to-transfect, protocols, mammalian call line, primary cells, sirna transfection, 165-2676, waveform independence, 165-2677, difficult to transfect, electroporate, 1652676 1652677, cell viability, plasmid dna, protocol optimization, LIT5582, rnai, transfecting buffers, universal reagent, nucleic acid delivery 5399 5399 Gene Pulser siRNA Electroporation References, Rev A /webroot/web/pdf/lsr/literature/Bulletin_5399.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Gene Pulser siRNA Electroporation References, Rev A Life Science 5399 gene silencing, citations, published citation, xcell system, cultured cell, LIT5399, reference, transformation, cells, silencing rna, excel, gene transfer 5445 Gene Pulser Xcell Electroporation System Flier, Rev A 5445 /webroot/web/pdf/lsr/literature/Bulletin_5445.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif No Gene Pulser Xcell Electroporation System Flier, Rev A 5445 165-2661, bacteria, cells, electroporator, exponential-decay, pulsetrac, 165-2667, 165-2669, 1652666, electroporators, 1652668, bacterial, dna, exponential decay, high voltage, high-voltage, prokaryote, pulse track, rna, 1652660, 1652662, LIT5445, nucleic acid, square wave, square-wave, transfection, 1652667, 1652669, fungi, mammalian cell, plants, shock-pod, 165-2660, 165-2662, eukaryote, genepulser, microbial, modular, shockpod, 165-2666, 1652661, acids, electro-poration, eukaryotic, fungal, plant, prokaryotic, transfect, 165-2668, eukaryotes, excel, prokaryotes, sirna 5598 Gene Pulser MXcell Electroporation System Flier, Rev B 5598 /webroot/web/pdf/lsr/literature/Bulletin_5598B.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif No Gene Pulser MXcell Electroporation System Flier, Rev B 5598 165-2674, 165-2676, 1652676, protocols, 165-2673, primary cells, trans fection, 1652673, delivery, transfection efficiency, buffer, instrument specifications, trans-fection, 165-2670, 1652670, cell, protocol, ShockPod cuvette chamber, optimization, optimize, 1652674 5634 Gene Pulser MXcell Electroporation System Brochure, Rev A 5634 /webroot/web/pdf/lsr/literature/Bulletin_5634.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif No Gene Pulser MXcell Electroporation System Brochure, Rev A 5634 plates, 5634, luciferase siRNA, plasmid delivery, LIT5634, preset protocols, well set, 5F2C, plate format, quick guide, HeLa, transfection conditions, Bulletin 5634, CHO cells, buffer, cell viability, square waveform, cell density, exponential decay waveform 1908 Electroporation Cuvette Flier, Rev B 1908 /webroot/web/pdf/lsr/literature/Bulletin_1908.pdf Literature PDF Product_Information_Sheets /webroot/web/images/general/icons/icon_pdf.gif No Electroporation Cuvette Flier, Rev B 1908 bulletin 1908, lit1908, bulletin 1908, transfection, trans-fection, genepulser, gene pulser, electroporation, electroporator, 165-2660, 165-2662, condition, protocol, shockpod, 1652661, efficiency, protocols, 1652081, 1652082, 1652083, 1652086, 1652088, 1652089, 1652091, 1652092, 1652093, 165-2661, pulsetrac, pulse trac, shock pod, 1652660, 1652662, 165-2660J1, 165-2661J1, 165-2662J1, 165-2666, 165-2666J1, 165-2667, 1652660J1, 1652661J1, 1652662J1, 1652666, 1652666J1, 1652667, 170-2503, 170-2503EDU, 170-2504, 170-2505, 1702503, 1702503EDU, 1702504, 1702505 5555 Protein Interaction Analysis Workflow Brochure, Rev B 5555 /webroot/web/pdf/lsr/literature/Bulletin_5555B.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif No Protein Interaction Analysis Workflow Brochure, Rev B 5555 biophysical characterization, Criterion cell, iQ5 multicolor detection, multi-plex, Profinity IMAC GST resins, ProteOn XPR36, Ready Prep 2 D, surface-enhanced laser desorption ionization, Tris-HCL gels, XPR-36, amplification, Model 491, multi-plexed, multi-plexing, RT-PCR, affinity chromatography, array, BioLogic DuoFlow F10 workstation, blotter, fractionation, micro-arrays, MicroRotofor, MP 5000, Power Pac, spectrophotometry, arrays, blotting, Gene Pulser/MicroPulser cuvettes, LC-MS/MS, MiniOpticon real-time PCR, Mutation, Versa Doc, assay, isoelectric focusing, MP-5000, profiling, Prote-On XPR 36, Bio-Logic Duo-Flow F-10, blot, iso-electric, microarrays, Mini-Opticon real time polymerase chain reaction, Profinia, ProteinChip SELDI, reverse transcription, reverse-transcription, sequential extraction, tagged purification, Versa-Doc, micro-array, Micro-Rotofor, MP5000, multi-color, multiplex, Power-Pac, PROTEAN IEF, ReadyPrep 2-D cleanup kit, silentfect lipid reagent for RNAi, surface enhanced desorption/ionization, antibody, assays, Experion automated electrophoresis system, Gene-Pulser Xcell total, high-throughput multiplexing, interface mapping, liquid chromatography-tandem mass spectrometry, microarray, Molecular Imager VersaDoc, multiplexed, PowerPac power supplies, PROTEON, Ready-Prep 2D, signal transduction pathway elucidation, transfectin 5554 Gene Silencing | RNAi Workflow Brochure, Rev B 5554 /webroot/web/pdf/lsr/literature/Bulletin_5554.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif No Gene Silencing | RNAi Workflow Brochure, Rev B 5554 1610449, 171-000201, 174-9974, 174-9975, 7007102, 732-6800, Aqua-pure, automated electrophoresis, blot, iso-electric, microarrays, microinjection, micropulser, mRNA quantification, multiplex suspension array technology, reverse transcription, reverse-transcription, RNA interference, silent-mer predesigned, small inhibitory RNAs, transfect, 1000, 161-0737, 163-2105, 1632130, 1652431, 1709780, 174-9970, 1749950, 700-7106, 7326370, BioOdyssey Calligrapher miniarrayer, micro-array, Mini-PROTEAN Tetra cell, multi-color, plasmid, ReadyPrep 2-D cleanup, ribonucleic acid, silentfect reagent, 165-4000, 1703350, 174-9960, 1749975, 1749977, 732-6370, 74-9950, amplification, Bio-Odyssey mini-arrayer, Laemmli buffer, multi-plexed, multi-plexing, multiplexing, PDS-1000 He, RT-PCR, spotting, transfected, vector-mediated, 165-2432, 170-3350, 171000201, 174-9961, 1749974, 1749976, 700-7102, 732-6890, Bio-Plex 200, biolistic particle delivery, Experion, Gene-Pulser Xcell total system, isoelectric focusing, micro-injection, micro-pulser, PDS-1000/He, Pure-Zol, validated siRNAs, 1610737, 165-2257, 1652088, 1652257, 1652432, 169-2000, 1692000, 170-9780, 174-9971, 1749971, blotter, cyclers, delivery optimization kit, IEF, qPCR, RT-qPCR, shRNA, 161-0449, 165-2431, 165-2660, 1703360, 1749961, 1749973, 7007106, aqua pure, arrayer, Bio Plex, chemically-mediated, genomics, Helios gun, iQ5 multicolor detection, Mini PROTEAN, multi-plex, nucleic, proteomics, Ready Prep 2 D, Real time polymerase chain reaction, Zol, 163-2130, 165-2088, 174-9972, 174-9977, 1749960, 1749970, 1749972, 7326800, 7326890, Aquapure isolation, arrays, BioPlex, chemically mediated, lipid transfection, LIT5554, PDS 1000/He, PureZol reagent, shRNAs, silentmer dicer-substrate siRNA duplexes, thermal cycler, 161-0490, 1610490, 1632105, 1652660, 1654000, 170-3360, 174-9973, 174-9976, 700-7104, 7007104, Aurum 96, electroporation, GenePulser cuvettes, microarray, multiplexed, Protein expression analysis, PROTEON, ready-prep 2D, real-time PCR, short hairpin RNAs, specific inhibition, thermal-cyclers, transfectin, western blotting 5924 Stem Cell Basics for Life Science Researchers Brochure, Rev A 5924 H /webroot/web/pdf/lsr/literature/Bulletin_5924A.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/icon_pdf.gif No Stem Cell Basics for Life Science Researchers Brochure, Rev A 5924 Bulletin 5924, embryonic differentiation, embryo, isolated pluripotent sc, development, multipotent, unipotent, terminally differentiated, blastocyst, mass of inner cells, ICM, es, history, hes, mes, human, mouse, lines, adult, somatic, imprinting, chimeric, programming, reprogramming, ips, isolation and maintenance, growth factors, media, medium, tissue culture, fibroblast, fibroblasts, feeder layer, hematopoietic, induced, ectoderm, mesoderm, endoderm, transfection, knock down, RNAi, viral-mediated delivery, lipid-mediated, electroporation, biolistic particle, retrovirus, genomic analysis, proteomic, expression, phenotype, karotyping, SNPs, SNP, PCR, microarray, profiling, profile, qPCR, array, flow cytometry, immunocytochemistry, RT-PCR, RT-qPCR, western blotting, Bio-Plex suspension array system, sso7d fusion protein technology, 170-8891, 170-8893, 170-8895, 170-8899, 172-5203, 170-8885, 172-5213, 172-5103, 172-5853, 172-5233, 170-8864, 172-5849, 172-5253, 172-5243, 172-5108, 172-5857, 165-2661, 165-2670, 165-2674, 165-2677, 165-2681, 165-2081, 165-2082, 165-2086, 165-2088, 165-2091, 165-2092, 170-8201, 170-8351, 170-8202, 170-8352, 170-8206, 170-8353, 170-8203, 170-8354, 170-8204, 170-8205, 170-8200, 1708891, 1708893, 1708895, 1708899, 1725203, 1708885, 1725213, 1725103, 1725853, 1725233, 1708864, 1725849, 1725253, 1725243, 1725108, 1725857, 1652661, 1652670, 1652674, 1652677, 1652681, 1652081, 1652082, 1652086, 1652088, 1652091, 1652092, 1708201, 1708351, 1708202, 1708352, 1708206, 1708353, 1708203, 1708354, 1708204, 1708205, 1708200 1349 Electroporation of T-Cell and Macrophage Cell Lines 1349 /webroot/web/pdf/lsr/literature/Bulletin_1349.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif No Electroporation of T-Cell and Macrophage Cell Lines 1349 lit1349, bulletin 1349, t-cell, macrophage, electroporation, gene pulser, gene transfer, dna transfer, 165-2660, 165-2661, 165-2662, 165-2666, 165-2667, 165-2668, 165-2669, 1652660, 1652661, 1652662, 1652666, 1652667, 1652668, 1652669, transfection, trans-fection, transformation, genepulser, electroporator, condition, protocol, shockpod, efficiency, protocols, 1652081, 1652082, 1652083, 1652086, 1652088, 1652089, 1652091, 1652092, 1652093, pulsetrac, pulse trac, shock pod, 165-2660J1, 165-2661J1, 165-2662J1, 165-2666J1, 1652660J1, 1652661J1, 1652662J1, 1652666J1, 170-2503, 170-2503EDU, 170-2504, 170-2505, 1702503, 1702503EDU, 1702504, 1702505 1365 Introducing Proteins Into Cells by Electroporation 1365 /webroot/web/pdf/lsr/literature/Bulletin_1365.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Introducing Proteins Into Cells by Electroporation 1365 lit1365, bulletin 1365, protein, transfer, cell, cellular, intracellular, electroporation, 165-2660, 165-2661, 165-2662, 165-2666, 165-2667, 165-2668, 165-2669, 1652660, 1652661, 1652662, 1652666, 1652667, 1652668, 1652669, transfection, trans-fection, transformation, genepulser, gene pulser, electroporator, condition, protocol, shockpod, efficiency, protocols, 1652081, 1652082, 1652083, 1652086, 1652088, 1652089, 1652091, 1652092, 1652093, pulsetrac, pulse trac, shock pod, 165-2660J1, 165-2661J1, 165-2662J1, 165-2666J1, 1652660J1, 1652661J1, 1652662J1, 1652666J1, 170-2503, 170-2503EDU, 170-2504, 170-2505, 1702503, 1702503EDU, 1702504, 1702505 1345 Electroporation of Primary Bone Marrow Cells 1345 /webroot/web/pdf/lsr/literature/Bulletin_1345.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif No Electroporation of Primary Bone Marrow Cells 1345 lit1345, bulletin 1345, bone, marrow, gene pulser, gene transfer, electroporation, 165-2660, 165-2661, 165-2662, 165-2666, 165-2667, 165-2668, 165-2669, 1652660, 1652661, 1652662, 1652666, 1652667, 1652668, 1652669, transfection, trans-fection, transformation, genepulser, electroporator, condition, protocol, shockpod, efficiency, protocols, 1652081, 1652082, 1652083, 1652086, 1652088, 1652089, 1652091, 1652092, 1652093, pulsetrac, pulse trac, shock pod, 165-2660J1, 165-2661J1, 165-2662J1, 165-2666J1, 1652660J1, 1652661J1, 1652662J1, 1652666J1, 170-2503, 170-2503EDU, 170-2504, 170-2505, 1702503, 1702503EDU, 1702504, 1702505 1355 Production of Hybridomas by Electrofusion 1355 /webroot/web/pdf/lsr/literature/Bulletin_1355.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif No Production of Hybridomas by Electrofusion 1355 lit1355, bulletin 1355, cell fusion, hybridoma, gene pulser, antigen, antibody, electrofusion, 165-2660, 165-2661, 165-2662, 165-2666, 165-2667, 165-2668, 165-2669, 1652660, 1652661, 1652662, 1652666, 1652667, 1652668, 1652669, transfection, trans-fection, transformation, genepulser, electroporation, electroporator, condition, protocol, shockpod, efficiency, protocols, 1652081, 1652082, 1652083, 1652086, 1652088, 1652089, 1652091, 1652092, 1652093, pulsetrac, pulse trac, shock pod, 165-2660J1, 165-2661J1, 165-2662J1, 165-2666J1, 1652660J1, 1652661J1, 1652662J1, 1652666J1, 170-2503, 170-2503EDU, 170-2504, 170-2505, 1702503, 1702503EDU, 1702504, 1702505 1365 1345 1349 5542 5858 The Gene Pulser MXcell Electroporation System Provides Reproducible Results in Electroporation Plates and Cuvettes With the Same Protocol, Rev A 5858 /webroot/web/pdf/lsr/literature/Bulletin_5858.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No The Gene Pulser MXcell Electroporation System Provides Reproducible Results in Electroporation Plates and Cuvettes With the Same Protocol, Rev A 5858 multiwell plate, waveforms, cho, chinese hamster ovary cells, conditions, cos-7, gene expression, viability, gfp, LIT5858, transfection efficiency, cell viability, condition, protocol, shockpod cuvette chamber, plates, protocols, waveform, exponential-decay pulse, square-wave 5641 The Gene Pulser MXcell Electroporation System Delivers Consistent Results Required for Optimizing Delivery Protocols, Rev A 5641 /webroot/web/pdf/lsr/literature/Bulletin_5641.pdf Literature PDF Protocols /webroot/web/images/general/icons/icon_pdf.gif No The Gene Pulser MXcell Electroporation System Delivers Consistent Results Required for Optimizing Delivery Protocols, Rev A 5641 gene silencing, LIT5641, luciferase activity, CHO, Bulletin 5641, 5641, HeLa cells, plasmid delivery, 5F2C, siRNA transfection 5603 Optimization of Electroporation Using Gene Pulser Electroporation Buffer and the Gene Pulser MXcell Electroporation System, Rev A 5603 /webroot/web/pdf/lsr/literature/Bulletin_5603.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Optimization of Electroporation Using Gene Pulser Electroporation Buffer and the Gene Pulser MXcell Electroporation System, Rev A 5603 silentMER (174-9974, luciferase expression plasmid, optimization studies, Exogenous nucleic acids, exponential-decay, HPF cells, LIT5603, 174-9975, siRNA, 174-9961), transfer, human primary fibroblasts, square-wave, transfection 5622 Optimization of Electroporation Conditions With the Gene Pulser MXcell Electroporation System, Rev A 5622 /webroot/web/pdf/lsr/literature/Bulletin_5622.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Optimization of Electroporation Conditions With the Gene Pulser MXcell Electroporation System, Rev A 5622 plasmid DNA, square waveform, LIT5622, Well set, transfection efficiency, difficult-to-transfect, primary, exponential, optimization protocol, siRNAs, interelectrode distance, square-wave 5686 Optimization of Electroporation Conditions for Jurkat Cells Using the Gene Pulser MXcell Electroporation System, Rev A 5686 /webroot/web/pdf/lsr/literature/Bulletin_5686.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Optimization of Electroporation Conditions for Jurkat Cells Using the Gene Pulser MXcell Electroporation System, Rev A 5686 5686, knockdown, Bulletin 5686, silencing, transfer, siLentMer, siRNA, plasmid DNA, luciferase activity assay, gene delivery, LIT5686, transfection 5687 Transfection of Mammalian Cells Using Preset Protocols on the Gene Pulser MXcell Electroporation System, Rev A 5687 /webroot/web/pdf/lsr/literature/Bulletin_5687.pdf Literature PDF Protocols /webroot/web/images/general/icons/icon_pdf.gif No Transfection of Mammalian Cells Using Preset Protocols on the Gene Pulser MXcell Electroporation System, Rev A 5687 short, trans-fection, efficiency, LIT5687, cho-k1, chok1, delivery, hela, sirna, expression, pcmviluc, plasmids, trans fection, condition, small interfering rna, cho, cell line, conditions, optimization, optimize, plasmid, viability 5720 Transfection of Neuroblastoma Cell Lines Using the Gene Pulser MXcell Electroporation System, Rev A 5720 /webroot/web/pdf/lsr/literature/Bulletin_5720.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Transfection of Neuroblastoma Cell Lines Using the Gene Pulser MXcell Electroporation System, Rev A 5720 delivery, LIT5720, siRNA, conditions, optimization, optimize, efficiencies, IMR-32, plasmid DNA, short, trans-fection, SK_N_SH, Cancer, pCMViLuc, plasmids, trans fection, efficiency, pCMVi-Luc, silencing, condition, gene expression analysis, small interfering RNA, neural cells 5704 Electroporation Conditions for Chinese Hamster Ovary Cells Using the Gene Pulser MXcell Electroporation System, Rev A 5704 /webroot/web/pdf/lsr/literature/Bulletin_5704.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Electroporation Conditions for Chinese Hamster Ovary Cells Using the Gene Pulser MXcell Electroporation System, Rev A 5704 efficiency, cell line, optimization, optimize, cho, cho-k1, chok1, LIT5704, trans-fection, expression, trans fection, transfection 5733 Transfection of Chinese Hamster Ovary-Derived DG44 Cells Using the Gene Pulser MXcell Electroporation System, Rev A 5733 /webroot/web/pdf/lsr/literature/Bulletin_5733.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Transfection of Chinese Hamster Ovary-Derived DG44 Cells Using the Gene Pulser MXcell Electroporation System, Rev A 5733 cell line, conditions, optimization, optimize, efficiency, pcmvi-luc, efficiencies, plasmid dna, short, trans-fection, delivery, sirna, cho, LIT5733, condition, small interfering rna, pcmviluc, plasmids, trans fection 5774 Delivery of siRNA by Electroporation Into Primary Human Neutrophils Using the Gene Pulser MXcell System, Rev A 5774 /webroot/web/pdf/lsr/literature/Bulletin_5774.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Delivery of siRNA by Electroporation Into Primary Human Neutrophils Using the Gene Pulser MXcell System, Rev A 5774 trans-fection, cell, cells, conditions, Fura-2 AM, inflammatory neutrophil activation, optimization, optimize, short interfering RNA, efficiency, flux, fMLP, RNAi, indicator, intracellular calcium concentration, trans fection, condition, LIT5774, mammalian, small, fluxes, transfection 5778 Electroporation Parameters for Transfection of HL-60 Leukocytic Cell Line With siRNA Using the Gene Pulser MXcell System, Rev B 5778 /webroot/web/pdf/lsr/literature/Bulletin_5778B.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Electroporation Parameters for Transfection of HL-60 Leukocytic Cell Line With siRNA Using the Gene Pulser MXcell System, Rev B 5778 inflammation, condition, inflammatory leukemic, leukocyte, LIT5778, primary cells, trans fection, trans-fection, RNAi, efficiency, neutrophil activation, RNA interference, conditions, leukocytes, optimization, optimize, short interfering RNA, mammalian, neutrophils, small 5823 Electroporation of Primary Murine Mast Cells Using the Gene Pulser MXcell Electroporation System, Rev A 5823 /webroot/web/pdf/lsr/literature/Bulletin_5823A.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Electroporation of Primary Murine Mast Cells Using the Gene Pulser MXcell Electroporation System, Rev A 5823 trans fection, conditions, green fluorescent protein, optimization, optimize, delivery, flow cytometry, transfection efficiency, cell viability, plasmid DNA, trans-fection, condition, protocol, reporter gene, protocols, transfect, GFP expression 5842 Optimization of Electroporation Conditions for Two Different Burkitt Lymphoma Cell Lines Using the Gene Pulser MXcell System, Rev B 5842 /webroot/web/pdf/lsr/literature/Bulletin_5842.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Optimization of Electroporation Conditions for Two Different Burkitt Lymphoma Cell Lines Using the Gene Pulser MXcell System, Rev B 5842 ramos, protocols, luciferase activity, namalwa cells, ldh level, protocol, exponential-decay condition, gene expression, viability, gfp, LIT5842, transfection efficiency, square-wave 5684 0108 0108 Simple and Rapid Optimization With Maximum Transfection Efficiency, Rev A /webroot/web/pdf/lsr/literature/Bulletin_0108.pdf Literature PDF Scientific_Posters /webroot/web/images/general/icons/icon_pdf.gif No Simple and Rapid Optimization With Maximum Transfection Efficiency, Rev A Life Science 0108 108, 0108, LIT0108, transfection, efficiency, mammalian cell, sirna 5860 Analysis of IL-4 Dependent Gene Expression in Namalwa Cells by siRNA Transfection: An Example of Pathway Analysis Using the Gene Pulser MXcell Electroporation System, Rev A 5860 /webroot/web/pdf/lsr/literature/Bulletin_5860A.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif No Analysis of IL-4 Dependent Gene Expression in Namalwa Cells by siRNA Transfection: An Example of Pathway Analysis Using the Gene Pulser MXcell Electroporation System, Rev A 5860 condition, ldh level, LIT5860, small, burkitt lymphoma cell line, knock down, trans-fection, burkitt’s, rnai, gapdh knockdown, tarc/ccl17, conditions, optimize, short interfering rna, stat6, primary cells, trans fection, ccl17 5904 Transfection of Mouse and Human Embryonic Stem Cells by Electroporation Using the Gene Pulser Mxcell&trade; System, Rev A 5904 /webroot/web/pdf/lsr/literature/Bulletin_5904.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/icon_pdf.gif Transfection of Mouse and Human Embryonic Stem Cells by Electroporation Using the Gene Pulser Mxcell&trade; System No Transfection of Mouse and Human Embryonic Stem Cells by Electroporation Using the Gene Pulser Mxcell&trade; System, Rev A 5904 transfection, trans-fection, genepulser, gene pulser, electroporation, electroporator, antibiotic resistance, buffer, dna delivery, expression, gene pulser ii, gene targeting, geneticin resistant, h9 es, mef, mouse, optimize, optimization, parameters, primary, trans-fection, trans fection, 165-2668, 1652668 2497 5443 Biolistic Particle Delivery Systems Brochure, Rev A 5443 /webroot/web/pdf/lsr/literature/Bulletin_5443.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif No Biolistic Particle Delivery Systems Brochure, Rev A 5443 165-2421, 165-2422, 1652422, cell-culture, chloroplasts, genes, Hepta adaptor, LIT5443, micro-carrier, micro-carriers, microparticle, plant, transfect, transfer, 165-2225, 165-2257, 165-2326, 165-2335, 1652225, 1652257, 1652278, 1652335, 1652432, bombardment, co-delivery, embryos, microcarrier, 1000/He, 165-2278, 1652326, 1652424, 1652431, bacteria, Helios Gene Gun System, mitochondrial, plasmid, transfecting, 165-2259, 165-2262, 165-2411, 165-2413, 165-2432, 1652413, bacterial, cotransformation, deoxyribonucleic, DNA, micro-particle, mitochondrion, PDS-1000/He, RNA, transform, yeast, 165-2258, 165-2336, 165-2412, 1652258, 1652336, cell culture, goldcoat, in vivo transformation, microcarriers, plasmids, 165-2431, 1652411, 1652418, acid-coated, bombard, chloroplast, micro-particles, rupture disks, 1652259, 1652421, co-transformation, codelivery, embryo, mitochondria, nucleic acid coated, ribonucleic, transfection, 165-2418, 165-2424, 1652262, 1652412, acceleration, animal, Heleos, helium pulse, microparticles, PDS 1000 He, pollen, trypsinized 5447 Biolistic PDS-1000/He System Flier, Rev A 5447 /webroot/web/pdf/lsr/literature/Bulletin_5447.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif No Biolistic PDS-1000/He System Flier, Rev A 5447 165-2225, 165-2257, 1652225, 1652257, acid, embryos, fungus, gold microparticle, particle bombardment, targets, chloroplasts, coridia, tissue culture, 165-2258, 1652258, biolistics, expression, fungi, bacteria, bio-listics, LIT5447, yeasts, algae, aspergillus nidulans, micro-particles, oganelles, shocking chamber, target, transforms, 165-2259, dna, helium, rna, yeast, 1652259, he, mitochondria, nucleic acid, pds1000, pds1000-he, transformation, fish embryo, gene transfer, microparticles, oganelle, pollen, tissues 5446 Helios Gene Gun System Flier, Rev A 5446 /webroot/web/pdf/lsr/literature/Bulletin_5446.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif No Helios Gene Gun System Flier, Rev A 5446 165-2431, 1652418, cartridge, geneshot, shot, target, 165-2421, 165-2422, 1652422, acids, barrel, LIT5446, micro-carrier, micro-carriers, plant cell culture, 1652424, 1652431, bacteria, bullet, cytomegalovirus, galactosidase, plasmid, projectiles, promoter, yeasts, 165-2413, 165-2432, 1652413, beta, biolistic, bullets, cmv, dna, microbe, rna, yeast, 165-2412, biolistics, expression, microcarriers, plants, plasmids, 165-2420, 1652421, cartridges, microparicles, nucleic acid, 1652432, beta-galacosidase, genome, gold microparticle, lacz, luciferase, microbes, microcarrier, projectile, targets, tissue, 165-2418, 165-2424, 1652412, 1652420, eukaryotes, gene transfer, helium pulse, luc, prokaryotes 2051 Transformation of Filamentous Fungi by Microprojectile Bombardment 2051 /webroot/web/pdf/lsr/literature/Bulletin_2051.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif No Transformation of Filamentous Fungi by Microprojectile Bombardment 2051 bulletin 2051, lit2051, transfection, trans-fection, microprojectile, mircoparticle, gold, tungsten, biolistic particle delivery, bombard, bombardment, transfection, trans-fection, protocol, helios, gene gun, pds-1000, pds-1000/he, fungi, fungus 2015 Sub-Micron Gold Particles Are Superior to Larger Particles for Efficient Biolistic Transformation of Organelles and Some Cell Types 2015 /webroot/web/pdf/lsr/literature/Bulletin_2015.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif No Sub-Micron Gold Particles Are Superior to Larger Particles for Efficient Biolistic Transformation of Organelles and Some Cell Types 2015 bulletin 2015, lit2015, transfection, trans-fection, microprojectile, mircoparticle, gold, tungsten, biolistic particle delivery, bombard, bombardment, transfection, trans-fection, protocol, helios, gene gun, pds-1000, pds-1000/he 2007 Bombardment-Mediated Transformation Methods for Barley 2007 /webroot/web/pdf/lsr/literature/Bulletin_2007.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif No Bombardment-Mediated Transformation Methods for Barley 2007 lit2007, bulletin 2007, transfection, trans-fection, microprojectile, mircoparticle, gold, tungsten, biolistic particle delivery, bombard, bombardment, transfection, trans-fection, protocol, helios, gene gun, pds-1000, pds-1000/he 2087 Biolistic Transfection of Organotypic Brain Slices and Dissociated Cells 2087 /webroot/web/pdf/lsr/literature/Bulletin_2087.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif No Biolistic Transfection of Organotypic Brain Slices and Dissociated Cells 2087 bulletin 2087, lit2087, transfection, trans-fection, microprojectile, mircoparticle, gold, tungsten, biolistic particle delivery, bombard, bombardment, transfection, trans-fection, protocol, helios, gene gun, pds-1000, pds-1000/he, brain, 1652258, 1652257, 165-2258, 165-2257 2658 2658 Single-Cell Complementation of Barley <i>mlo</i> Mutants Using a PDS-1000/He Hepta System /webroot/web/pdf/lsr/literature/Bulletin_2658.pdf Literature PDF Application Notes /webroot/web/images/general/icons/icon_pdf.gif No Single-Cell Complementation of Barley <i>mlo</i> Mutants Using a PDS-1000/He Hepta System Life Science 2658 PDS1000He, PDS1000, bulletin 2658, LIT2658 2453 2453 Optimization of Gene Delivery Into Arabidopsis, Tobacco, and Birch Using the Helios Gene Gun System /webroot/web/pdf/lsr/literature/Bulletin_2453.pdf Literature PDF Application Notes /webroot/web/images/general/icons/icon_pdf.gif No Optimization of Gene Delivery Into Arabidopsis, Tobacco, and Birch Using the Helios Gene Gun System Life Science 2453 bombard, biolistic particle delivery, microprojectile, transformation, LIT2453 1688 Optimization of Biolistic<sup>&reg;</sup> Transformation Using the Helium-Driven PDS-1000/He System 1688 /webroot/web/pdf/lsr/literature/Bulletin_1688.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif No Optimization of Biolistic Transformation Using the Helium-Driven PDS-1000/He System 1688 lit1688, bulletin 1688, biolistic, particle delivery, gene delivery, gene transfer, 165-2257, 165-2258, 165-2225, 165-2259, 1652257, 1652258, 1652225, 1652259 1689 Comparison of Performance Characteristics of Different Biolistic<sup>&reg;</sup> Devices 1689 /webroot/web/pdf/lsr/literature/Bulletin_1689.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif No Comparison of Performance Characteristics of Different Biolistic<sup>&reg;<sup> Devices 1689 lit1689, bulletin 1689, bombard, pds-1000/he, biolistic, particle delivery, 165-2411, 165-2431, 165-2451, 165-2257, 165-2258, 165-2225, 165-2259, 1652411, 1652431, 1652451, 1652257, 1652258, 1652225, 1652259 2433 2433 Transformation of Nematodes With the Helios Gene Gun /webroot/web/pdf/lsr/literature/Bulletin_2433.pdf Literature PDF Application Notes /webroot/web/images/general/icons/icon_pdf.gif No Transformation of Nematodes With the Helios Gene Gun Life Science 2433 biolistic particle delivery, bombard, bulletin 2433, gene transfer, LIT2433 2552 2552 The Gene Gun: Current Applications in Cutaneous Gene Therapy, Rev A /webroot/web/pdf/lsr/literature/Bulletin_2552.pdf Literature PDF Application Notes /webroot/web/images/general/icons/icon_pdf.gif No The Gene Gun: Current Applications in Cutaneous Gene Therapy, Rev A Life Science 2552 vaccination, biolistic particle delivery, helios biolistics, LIT2552, bulletin 2552, helios 2453 2531 Inoculation of Viral RNA and cDNA to Potato and Tobacco Plants Using the Helios Gene Gun 2531 /webroot/web/pdf/lsr/literature/Bulletin_2531.pdf Literature PDF /webroot/web/images/general/icons/icon_pdf.gif No Inoculation of Viral RNA and cDNA to Potato and Tobacco Plants Using the Helios Gene Gun 2531 transfer, bombardment, biolistic particle delivery, LIT2531, transformation, bulletin 2531 2410 Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios<sup>&reg;</sup> Gene Gun-Mediated Particle Bombardment, Rev B 2410 /webroot/web/pdf/lsr/literature/Bulletin_2410.pdf Literature PDF Manuals_and_Quick_Guides /webroot/web/images/general/icons/icon_pdf.gif Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios Gene Gun-Mediated Particle Bombardment No Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios Gene Gun-Mediated Particle Bombardment 2410 helios, helium, biolistic, biolistics, gene gun, projectile, microprojectile, microparticle, transfection, transformation, 165-2411, 165-2431, 165-2451, 1652411, 1652431, 1652451 2726 Delivery of pCMV-S DNA Using the Helios&reg; Gene Gun System Is Superior to Intramuscular Injection in Balb/c Mice 2726 /webroot/web/pdf/lsr/literature/Bulletin_2726.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif Delivery of pCMV-S DNA Using the Helios&reg; Gene Gun System Is Superior to Intramuscular Injection in Balb/c Mice No Delivery of pCMV-S DNA Using the Helios&reg; Gene Gun System Is Superior to Intramuscular Injection in Balb/c Mice 2726 helios, gene gun, transfection, gene transfer, biolistic, particle delivery, bulletin 2726, lit2726, 125-0224, 1250224 1689 2768 2768 Biolistic Gene Transfer to Generate Transgenic Schistosomes, Rev A /webroot/web/pdf/lsr/literature/Bulletin_2768.pdf Literature PDF Application Notes /webroot/web/images/general/icons/icon_pdf.gif No Biolistic Gene Transfer to Generate Transgenic Schistosomes, Rev A Life Science 2768 bulletin 2768, gene gun, LIT2768 Life Science Research/Products/Transfection/Electroporation/Gene Pulser Xcell Electroporation Systems ->MT::b1a35eb3-d55c-47b3-aaf3-95e4d1d85848##Life Science Research/Products/Transfection/Electroporation/Gene Pulser MXcell Electroporation System ->MT::fae0f825-da45-4b02-aa33-73781dda6171##Life Science Research/Products/Transfection/Electroporation/MicroPulser Electroporator ->MT::83527990-34fb-4b33-b955-ca53b57bf8b9##Life Science Research/Products/Transfection/Biolistic Particle Delivery Systems/Helios Gene Gun Systems ->MT::42e9d6be-369a-49f8-8fbb-281a0fea6df8##Life Science Research/Products/Transfection/Biolistic Particle Delivery Systems/PDS-1000 | He and Hepta Systems ->MT::1730e08d-f43a-46ea-b7f3-7b35c04c36eb## Life Science Research/Solutions/Technologies/qPCR|Real-Time PCR ->MTS::LUSO4W8UU##Life Science Research/Solutions/Technologies/Cell Counting Methods ->MTS::LUSOLB470##Life Science Research/Solutions/Technologies/Imaging and Analysis ->MTS::LUSQC6MNI##Life Science Research/Solutions/Technologies/Transfection ->MTS::LUSONCB9O## Eddie C Instrument-Based Transfection Methods 12/28/11 04:27 PM 12/28/21 04:37 PM AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA en LSR /LSR/Technologies/Transfection N 0 Introduction to Transfection /en-us/applications-technologies/applications-technologies/instrument-based-transfection-methods?ID=LUSONCB9O

Transfection of cells can be accomplished by various methods, including chemical, biological, and instrument-based. This section provides an overview of the different instrument-based transfection methods available, discusses how they work, and describes their pros and cons.

Related Topics: Chemical- and Viral-Based Transfection Methods, Posttransfection Analysis of Cells and Cell Counting Methods.

 

Overview of Instrument-Based Transfection Methods

Transfection can be accomplished using chemical, biological, or physical methods. Common methods include electroporation, the use of virus vectors, lipofection, and biolistics. Many types of genetic material, including plasmid DNA, siRNA, proteins, dyes, and antibodies may be transfected using any of these methods. However, no single method can be applied to all types of cells; transfection efficiencies and cytotoxicity may vary dramatically and depend on the method, cell type being utilized, and types of experiments being performed. Therefore, to obtain high efficiencies, all relevant factors should be considered in planning and selecting the appropriate transfection method.

Method Function Pros Cons Cells Products
Electroporation Nucleic acids or other molecules are introduced into cells by creating transient pores in the plasma membrane using an electric pulse Nonchemical method that doesn't seem to alter the biological structure or function of the target cells

Easy to perform

High efficiency

Can be applied to a wide range of cell types
Cell mortality (if using suboptimal conditions) Eukaryotic cells (primary, stem cells)

Prokaryotic cells (bacteria, yeast)

Plant protoplasts
Gene Pulser Xcell electroporation system

Gene Pulser MXcell electroporation system

MicroPulser electroporator
Biolistic particle delivery Delivery of nucleic acids into cells via high-velocity nucleic acid-coated microparticles Simple, rapid, versatile technique

Targeted intracellular gene delivery

Cell type independent

Uses small amounts of DNA

Delivers single or multiple genes

No carrier DNA needed

Can deliver large DNA fragments

No extraneous genes or proteins delivered

Requires little manipulation of cells

High reproducibility
Generally lower efficiency compared to electroporation or viral or lipid mediated transfection

Limited bacterial transfection data

Requires the preparation of microparticles

Instrument cost

Requires purchase agreement
Plant

Primary cells

Tissue

In vivo applications
Helios® gene gun system

PDS-1000/He and Hepta systems
Microinjection Direct injection of naked DNA Can be used for many animals Laborious (one cell at a time)

Technically demanding and costly
Eukaryotic cells  
Laserfection/
optoinjection
Uses laser light to transiently permeabilize a large number of cells in a very short time Very efficient

Works with many cell types

Few cell manipulations needed

Requires cell to be attached

Expensive laser equipment required
Attached cells  
 

Electroporation

  • 1. Electroporation exposes a cell to a high-intensity electric field that temporarily destabilizes the membrane
  • 2. During this time the membrane is highly permeable to exogenous molecules present in the surrounding media
  • 3. DNA then moves into the cell through these holes
  • 4. When the field is turned off, the pores in the membrane reseal, enclosing the DNA inside

Electroporation of cells.

 

Biolistic Particle Delivery

Biolistics is the delivery of nucleic acids into cells by firing nucleic acid-coated microparticles into them.

Helios Gene Gun

  • For in situ, in vivo and in vitro transformations
  • Applications for animals, plants, cell culture, nematodes, yeast and bacteria
  • Pressure range 100–600 psi enables fine-tuning of penetration
  • Highly portable can be used in the field
  • Small target area for accurate targeting

Biolistic particle delivery workflow.

PDS-1000/He Biolistic Particle Delivery System

  • For in vitro, ex vivo (and in vivo for some plants and microbes)
  • Applications for animal cell and organ culture, plant cell culture and explants, pollen, insects, algae, fungi and bacteria
  • Pressure range 450–2200 psi gives flexibility and penetration — ideal for plant applications
  • Large target area — more cells can be transformed

Procedure

  1. DNA-coated microcarriers (thin plastic disk) are spread over the central area of that disk using a pipette tip.
  2. Disk loaded with the DNA-coated particles is placed into a holder inside the PDS-1000 system.
  3. The system uses high pressure helium, released by a rupture disk, and a partial vacuum, to propel the macrocarrier sheet loaded with DNA-coated gold macrocarriers toward the target cells.
  4. Macrocarrier is halted after a short distance by a stopping screen.
  5. DNA-coated particles continue traveling toward the target to penetrate the cells.
  6. Sample chamber is subjected to a partial vacuum, from 15 to 29 in. of mercury, depending on the target cells.

 

Workflow for delivery using PDS-1000/He system.

 

Microinjection

  • Direct injection of naked DNA
  • Laborious (one cell at a time)
  • Technically demanding and costly
  • Can be used for many animals

Microinjection of particles.

 

Laserfection/Optoinjection

  • This procedure uses laser light to transiently permeabilize a large number of cells in a very short time
  • Various substances, including ions, small molecules, dextrans, short interfering RNAs (siRNAs), plasmids, proteins, and semiconductor nanocrystals can be efficiently optoinjected into numerous cell types

Workflow for laserfection.

 

Transfection Protocol Library

The transfection protocol online library contains protocols obtained from the literature, developed by Bio-Rad scientists, or submitted by scientists like you. Browse protocols to view our library and find your starting point or submit a protocol by clicking the proper technology.

 

Further Reading

Belyansteva IA (2009). Helios Gene Gun-mediated transfection of the inner ear sensory epithelium. Methods Mol Biol 493, 103–123. PMID: 18839344

Benediktsson AM et al. (2005). Ballistic labeling and dynamic imaging of astrocytes in organotypic hippocampal slice cultures. J Neurosci Methods 141, 41–53. PMID: 15585287

Eizema K et al. (2000). Endothelin-1 responsiveness of a 1.4 kb phospholamban promoter fragment in rat cardiomyocytes transfected by the gene gun. J Mol Cell Cardiol 32, 311–321. PMID: 10722806

Fujiki R et al. (2009). GlcNAcylation of a histone methyltransferase in retinoic-acid-induced granulopoiesis. Nature 459, 455–459. PMID: 19377461

Gildea JJ et al. (2009). Caveolin-1 and dopamine-mediated internalization of NaKATPase in human renal proximal tubule cells. Hypertension 54, 1070–1076. PMID: 19752292

Helledie T et al. (2008). A simple and reliable electroporation method for human bone marrow mesenchymal stem cells. Stem Cells Dev 17, 837–848. PMID: 18752428

Hockemeyer D et al. (2009). Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases. Nat Biotechnol 27, 851–857. PMID: 19680244

Huang B et al. (2008). RNA interference-mediated in vivo silencing of fas ligand as a strategy for the enhancement of DNA vaccine potency. Hum Gene Ther 19, 763–773. PMID: 18627219

Shimamura K et al. (2007). Generation of secondary small interfering RNA in cell-autonomous and non-cell autonomous RNA silencing in tobacco. Plant Mol Biol 63, 803–813. PMID: 17225952

Su L et al. (2009). Neural stem cell differentiation is mediated by integrin beta4 in vitro. Int J Biochem Cell Biol 41, 916–924. PMID: 18834954

Tseng CN et al. (2013). A method to identify RNA A-to-I editing targets using I-specific cleavage and exon array analysis. Mol Cell Probes 7, 38–45. PMID: 22960667

Wirth MJ and Wahle P (2003). Biolistic transfection of organotypic cultures of rat visual cortex using a handheld device. J Neurosci Methods 125, 45–54. PMID: 12763229

Zhang G and Selzer ME (2001). In vivo transfection of lamprey brain neurons by gene gun delivery of DNA. Exp Neurol 167, 304–311. PMID: 11161618

 

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This tutorial highlights the main components and features of the Gene Pulser Xcell system. It provides information about system installation and the setup of electroporation experiments, including important troubleshooting tips and answers to frequently asked questions. Ordering information for system components and accessories is also provided.
 
Literature
Number Description Download
5448 MicroPulser Electroporator Flier, Rev A Click to download
5542 Electroporation Systems Brochure, Rev A Click to download
5582 Gene Pulser Electroporation Buffer Product Information Sheet, Rev A Click to download
5399 Gene Pulser siRNA Electroporation References, Rev A Click to download
5445 Gene Pulser Xcell Electroporation System Flier, Rev A Click to download
5598 Gene Pulser MXcell Electroporation System Flier, Rev B Click to download
5634 Gene Pulser MXcell Electroporation System Brochure, Rev A Click to download
1908 Electroporation Cuvette Flier, Rev B Click to download
5555 Protein Interaction Analysis Workflow Brochure, Rev B Click to download
5554 Gene Silencing | RNAi Workflow Brochure, Rev B Click to download
5924 Stem Cell Basics for Life Science Researchers Brochure, Rev A Click to download
1349 Electroporation of T-Cell and Macrophage Cell Lines Click to download
1365 Introducing Proteins Into Cells by Electroporation Click to download
1345 Electroporation of Primary Bone Marrow Cells Click to download
1355 Production of Hybridomas by Electrofusion Click to download
5858 The Gene Pulser MXcell Electroporation System Provides Reproducible Results in Electroporation Plates and Cuvettes With the Same Protocol, Rev A Click to download
5641 The Gene Pulser MXcell Electroporation System Delivers Consistent Results Required for Optimizing Delivery Protocols, Rev A Click to download
5603 Optimization of Electroporation Using Gene Pulser Electroporation Buffer and the Gene Pulser MXcell Electroporation System, Rev A Click to download
5622 Optimization of Electroporation Conditions With the Gene Pulser MXcell Electroporation System, Rev A Click to download
5686 Optimization of Electroporation Conditions for Jurkat Cells Using the Gene Pulser MXcell Electroporation System, Rev A Click to download
5687 Transfection of Mammalian Cells Using Preset Protocols on the Gene Pulser MXcell Electroporation System, Rev A Click to download
5720 Transfection of Neuroblastoma Cell Lines Using the Gene Pulser MXcell Electroporation System, Rev A Click to download
5704 Electroporation Conditions for Chinese Hamster Ovary Cells Using the Gene Pulser MXcell Electroporation System, Rev A Click to download
5733 Transfection of Chinese Hamster Ovary-Derived DG44 Cells Using the Gene Pulser MXcell Electroporation System, Rev A Click to download
5774 Delivery of siRNA by Electroporation Into Primary Human Neutrophils Using the Gene Pulser MXcell System, Rev A Click to download
5778 Electroporation Parameters for Transfection of HL-60 Leukocytic Cell Line With siRNA Using the Gene Pulser MXcell System, Rev B Click to download
5823 Electroporation of Primary Murine Mast Cells Using the Gene Pulser MXcell Electroporation System, Rev A Click to download
5842 Optimization of Electroporation Conditions for Two Different Burkitt Lymphoma Cell Lines Using the Gene Pulser MXcell System, Rev B Click to download
0108 Simple and Rapid Optimization With Maximum Transfection Efficiency, Rev A Click to download
5860 Analysis of IL-4 Dependent Gene Expression in Namalwa Cells by siRNA Transfection: An Example of Pathway Analysis Using the Gene Pulser MXcell Electroporation System, Rev A Click to download
5904 Transfection of Mouse and Human Embryonic Stem Cells by Electroporation Using the Gene Pulser Mxcell&trade; System, Rev A Click to download
5443 Biolistic Particle Delivery Systems Brochure, Rev A Click to download
5447 Biolistic PDS-1000/He System Flier, Rev A Click to download
5446 Helios Gene Gun System Flier, Rev A Click to download
2051 Transformation of Filamentous Fungi by Microprojectile Bombardment Click to download
2015 Sub-Micron Gold Particles Are Superior to Larger Particles for Efficient Biolistic Transformation of Organelles and Some Cell Types Click to download
2007 Bombardment-Mediated Transformation Methods for Barley Click to download
2087 Biolistic Transfection of Organotypic Brain Slices and Dissociated Cells Click to download
2658 Single-Cell Complementation of Barley <i>mlo</i> Mutants Using a PDS-1000/He Hepta System Click to download
2453 Optimization of Gene Delivery Into Arabidopsis, Tobacco, and Birch Using the Helios Gene Gun System Click to download
1688 Optimization of Biolistic<sup>&reg;</sup> Transformation Using the Helium-Driven PDS-1000/He System Click to download
1689 Comparison of Performance Characteristics of Different Biolistic<sup>&reg;</sup> Devices Click to download
2433 Transformation of Nematodes With the Helios Gene Gun Click to download
2552 The Gene Gun: Current Applications in Cutaneous Gene Therapy, Rev A Click to download
2531 Inoculation of Viral RNA and cDNA to Potato and Tobacco Plants Using the Helios Gene Gun Click to download
2410 Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios<sup>&reg;</sup> Gene Gun-Mediated Particle Bombardment, Rev B Click to download
2726 Delivery of pCMV-S DNA Using the Helios&reg; Gene Gun System Is Superior to Intramuscular Injection in Balb/c Mice Click to download
2768 Biolistic Gene Transfer to Generate Transgenic Schistosomes, Rev A Click to download
6176 Electroporation Systems Overview Click to download
6177 Biolistic Particle Delivery Systems Click to download
6178 Recommended Biolistic System by Cell Types Click to download
6179 Lipid Transfection Reagents Selection Guide Click to download
 
 
LUSONV30E [referer] = [http://www.bio-rad.com/en-us/product/cell-sorting/..product/cell-sorting/gene-pulser-xcell-electroporation-systems?ID=b1a35eb3-d55c-47b3-aaf3-95e4d1d85848] [x-forwarded-proto] = [http] [x-forwarded-port] = [80] [x-forwarded-for] = [5.62.35.15, 10.232.2.13] [dnt] = [1] [accept] = [text/html, application/xml;q=0.9, application/xhtml+xml, image/png, image/webp, image/jpeg, image/gif, image/x-xbitmap, */*;q=0.1] [seourl] = [/en-us/applications-technologies/transfection-transduction-stem-cells] [x-amzn-trace-id] = [Root=1-5b506dbf-534a17ac1def93b7a8c13b13] [x-forwarded-server] = [lsds-prod-s.br.aws-livesite.io] [x-forwarded-host] = [www.bio-rad.com] [x-query-string] = [ID=LUSONV30E] [host] = [10.232.16.28:1776] [x-request-uri] = [/en-us/applications-technologies/transfection-transduction-stem-cells] [connection] = [Keep-Alive] [accept-encoding] = [gzip, deflate] [user-agent] = [Mozilla/5.0 (Windows NT 6.3; WOW64; Trident/7.0; rv:11.0) like Gecko] AppTech/AppTechDetails pageStyleKey internet/solutions_sub applications-technologies/transfection-transduction-stem-cells LSR LUSONV30E Instrument based Instrument-Based Transfection Methods /webroot/web/html/lsr/solutions/technologies/transfection <p>Transfection of cells can be accomplished by various methods, including chemical, biological, and instrument-based. This section provides an overview of the different instrument-based transfection methods available, discusses how they work, and describes their pros and cons.</p> <p><strong>Related Topics</strong>: <a href="/evportal/destination/solutions?catID=LUSOOP49">Chemical- and Viral-Based Transfection Methods</a>, <a href="/evportal/destination/solutions?catID=LUSOPS84">Posttransfection Analysis of Cells</a> and <a href="/evportal/destination/solutions?catID=LUSOLB470">Cell Counting Methods</a>.</p> Overview of Instrument-Based Transfection Methods <p>Transfection can be accomplished using chemical, biological, or physical methods. Common methods include electroporation, the use of virus vectors, lipofection, and biolistics. Many types of genetic material, including plasmid DNA, siRNA, proteins, dyes, and antibodies may be transfected using any of these methods. However, no single method can be applied to all types of cells; transfection efficiencies and cytotoxicity may vary dramatically and depend on the method, cell type being utilized, and types of experiments being performed. Therefore, to obtain high efficiencies, all relevant factors should be considered in planning and selecting the appropriate transfection method.</p> <table class="pd_table pd_gridlines txttop" border="0"> <tbody> <tr class="pd_colorbackground"> <td><strong>Method</strong></td> <td><strong>Function</strong></td> <td><strong>Pros</strong></td> <td><strong>Cons</strong></td> <td><strong>Cells</strong></td> <td><strong>Products</strong></td> </tr> <tr> <td valign="top">Electroporation</td> <td valign="top">Nucleic acids or other molecules are introduced into cells by creating transient pores in the plasma membrane using an electric pulse</td> <td valign="top">Nonchemical method that doesn't seem to alter the biological structure or function of the target cells<br /><br /> Easy to perform<br /><br /> High efficiency<br /><br /> Can be applied to a wide range of cell types</td> <td valign="top">Cell mortality (if using suboptimal conditions)</td> <td valign="top">Eukaryotic cells (primary, stem cells)<br /><br /> Prokaryotic cells (bacteria, yeast)<br /><br />Plant protoplasts</td> <td valign="top"><a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=b1a35eb3-d55c-47b3-aaf3-95e4d1d85848">Gene Pulser Xcell<sup>&trade;</sup> electroporation system</a><br /> <br /> <a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=fae0f825-da45-4b02-aa33-73781dda6171">Gene Pulser MXcell<sup>&trade;</sup> electroporation system</a><br /> <br /> <a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=83527990-34fb-4b33-b955-ca53b57bf8b9">MicroPulser<sup>&trade;</sup> electroporator</a></td> </tr> <tr> <td valign="top">Biolistic particle delivery</td> <td valign="top">Delivery of nucleic acids into cells via high-velocity nucleic acid-coated microparticles</td> <td valign="top">Simple, rapid, versatile technique<br /><br /> Targeted intracellular gene delivery<br /><br /> Cell type independent<br /><br /> Uses small amounts of DNA<br /><br /> Delivers single or multiple genes<br /><br /> No carrier DNA needed<br /><br /> Can deliver large DNA fragments<br /><br /> No extraneous genes or proteins delivered<br /><br /> Requires little manipulation of cells<br /><br /> High reproducibility</td> <td valign="top">Generally lower efficiency compared to electroporation or viral or lipid mediated transfection<br /><br /> Limited bacterial transfection data<br /><br /> Requires the preparation of microparticles<br /><br /> Instrument cost<br /><br /> Requires purchase agreement</td> <td valign="top">Plant<br /><br /> Primary cells<br /><br /> Tissue<br /><br /> In vivo applications</td> <td valign="top"><a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=42e9d6be-369a-49f8-8fbb-281a0fea6df8">Helios<sup>&reg;</sup> gene gun system</a><br /> <br /> <a href="http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=1730e08d-f43a-46ea-b7f3-7b35c04c36eb">PDS-1000/He and Hepta<sup>&trade;</sup> systems</a></td> </tr> <tr> <td valign="top">Microinjection</td> <td valign="top">Direct injection of naked DNA</td> <td valign="top">Can be used for many animals</td> <td valign="top">Laborious (one cell at a time)<br /><br /> Technically demanding and costly</td> <td valign="top">Eukaryotic cells</td> <td valign="top">&nbsp;</td> </tr> <tr> <td valign="top">Laserfection/<br /> optoinjection</td> <td valign="top">Uses laser light to transiently permeabilize a large number of cells in a very short time</td> <td valign="top">Very efficient<br /><br /> Works with many cell types<br /><br /> Few cell manipulations needed<br /><br /></td> <td valign="top">Requires cell to be attached<br /><br /> Expensive laser equipment required</td> <td valign="top">Attached cells</td> <td valign="top">&nbsp;</td> </tr> </tbody> </table> <div class="top"><a href="#helptop">Back to Top</a></div> Electroporation <p><img src="/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img1.jpg" alt="" width="245" height="624" align="left" /></p> <ul style="list-style-type: none;"> <li style="margin-bottom: 125px;">1. Electroporation exposes a cell to a high-intensity electric field that temporarily destabilizes the membrane</li> <li style="margin-bottom: 135px;">2. During this time the membrane is highly permeable to exogenous molecules present in the surrounding media</li> <li style="margin-bottom: 145px;">3. DNA then moves into the cell through these holes</li> <li>4. When the field is turned off, the pores in the membrane reseal, enclosing the DNA inside</li> </ul> <p class="caption" style="clear: both;"><strong>Electroporation of cells.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> Biolistic Particle Delivery <p>Biolistics is the delivery of nucleic acids into cells by firing nucleic acid-coated microparticles into them.</p> <p><strong>Helios Gene Gun</strong></p> <ul> <li>For in situ, in vivo and in vitro transformations</li> <li>Applications for animals, plants, cell culture, nematodes, yeast and bacteria</li> <li>Pressure range 100&ndash;600 psi enables fine-tuning of penetration</li> <li>Highly portable can be used in the field</li> <li>Small target area for accurate targeting</li> </ul> <p><img src="/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img2.jpg" alt="" width="560" height="270" /></p> <p class="caption"><strong>Biolistic particle delivery workflow.</strong></p> <p><strong>PDS-1000/He Biolistic Particle Delivery System</strong></p> <ul> <li>For in vitro, ex vivo (and in vivo for some plants and microbes)</li> <li>Applications for animal cell and organ culture, plant cell culture and explants, pollen, insects, algae, fungi and bacteria</li> <li>Pressure range 450&ndash;2200 psi gives flexibility and penetration &mdash; ideal for plant applications</li> <li>Large target area &mdash; more cells can be transformed</li> </ul> <p><strong>Procedure</strong></p> <ol> <li>DNA-coated microcarriers (thin plastic disk) are spread over the central area of that disk using a pipette tip.</li> <li>Disk loaded with the DNA-coated particles is placed into a holder inside the PDS-1000 system.</li> <li>The system uses high pressure helium, released by a rupture disk, and a partial vacuum, to propel the macrocarrier sheet loaded with DNA-coated gold macrocarriers toward the target cells.</li> <li>Macrocarrier is halted after a short distance by a stopping screen.</li> <li>DNA-coated particles continue traveling toward the target to penetrate the cells.</li> <li>Sample chamber is subjected to a partial vacuum, from 15 to 29 in. of mercury, depending on the target cells.</li> </ol> <p>&nbsp;</p> <p><img src="/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img3.jpg" alt="" width="560" height="224" /></p> <p class="caption"><strong>Workflow for delivery using PDS-1000/He system.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> Microinjection <ul> <li>Direct injection of naked DNA</li> <li>Laborious (one cell at a time)</li> <li>Technically demanding and costly</li> <li>Can be used for many animals</li> </ul> <p><img src="http://qaevn.bio-rad.com/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img4.jpg" alt="" width="532" height="172" /></p> <p class="caption"><strong>Microinjection of particles.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> Laserfection/Optoinjection <ul> <li>This procedure uses laser light to transiently permeabilize a large number of cells in a very short time</li> <li>Various substances, including ions, small molecules, dextrans, short interfering RNAs (siRNAs), plasmids, proteins, and semiconductor nanocrystals can be efficiently optoinjected into numerous cell types</li> </ul> <p><img src="http://qaevn.bio-rad.com/webroot/web/images/lsr/solutions/technologies/gene_expression/pcr/technology_detail/gxt41_img5.jpg" alt="" width="532" height="144" /></p> <p class="caption"><strong>Workflow for laserfection.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> Transfection Protocol Library <p> <script type="text/javascript"><!-- // popupwin function popIt(url,w,h,r){ var mydate = new Date(); wname=''+mydate.getMonth()+mydate.getDate()+mydate.getHours()+mydate.getMinutes()+mydate.getSeconds(); if (w && h && r) { popwin = window.open(url,"popwin"+wname,"height="+h+",width="+w+",status=1,scrollbars=1,location=1,menubar=1,resizable"); } else { popwin = window.open(url,"popwin"+wname,"height="+h+",width="+w+",status=1,scrollbars=1,location=0,resizable"); } } // --></script> </p> <p>The transfection protocol online library contains protocols obtained from the literature, developed by Bio-Rad scientists, or submitted by scientists like you. <a onclick="popIt('/genetransferprotocols',1300,700);return false" href="#" target="_blank">Browse protocols</a> to view our library and find your starting point or submit a protocol by clicking the proper technology.</p> <div class="top"><a href="#helptop">Back to Top</a></div> Further Reading <p>Belyansteva IA (2009). Helios Gene Gun-mediated transfection of the inner ear sensory epithelium. Methods Mol Biol 493, 103&ndash;123. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/18839344" target="_blank">18839344</a></p> <p>Benediktsson AM et al. (2005). Ballistic labeling and dynamic imaging of astrocytes in organotypic hippocampal slice cultures. J Neurosci Methods 141, 41&ndash;53. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/15585287" target="_blank">15585287</a></p> <p>Eizema K et al. (2000). Endothelin-1 responsiveness of a 1.4 kb phospholamban promoter fragment in rat cardiomyocytes transfected by the gene gun. J Mol Cell Cardiol 32, 311&ndash;321. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/10722806" target="_blank">10722806</a></p> <p>Fujiki R et al. (2009). GlcNAcylation of a histone methyltransferase in retinoic-acid-induced granulopoiesis. Nature 459, 455&ndash;459. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/19377461" target="_blank">19377461</a></p> <p>Gildea JJ et al. (2009). Caveolin-1 and dopamine-mediated internalization of NaKATPase in human renal proximal tubule cells. Hypertension 54, 1070&ndash;1076. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/19752292" target="_blank">19752292</a></p> <p>Helledie T et al. (2008). A simple and reliable electroporation method for human bone marrow mesenchymal stem cells. Stem Cells Dev 17, 837&ndash;848. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/18752428" target="_blank">18752428</a></p> <p>Hockemeyer D et al. (2009). Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases. Nat Biotechnol 27, 851&ndash;857. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/19680244" target="_blank">19680244</a></p> <p>Huang B et al. (2008). RNA interference-mediated in vivo silencing of fas ligand as a strategy for the enhancement of DNA vaccine potency. Hum Gene Ther 19, 763&ndash;773. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/18627219" target="_blank">18627219</a></p> <p>Shimamura K et al. (2007). Generation of secondary small interfering RNA in cell-autonomous and non-cell autonomous RNA silencing in tobacco. Plant Mol Biol 63, 803&ndash;813. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/17225952" target="_blank">17225952</a></p> <p>Su L et al. (2009). Neural stem cell differentiation is mediated by integrin beta4 in vitro. Int J Biochem Cell Biol 41, 916&ndash;924. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/18834954" target="_blank">18834954</a></p> <p>Tseng CN et al. (2013). A method to identify RNA A-to-I editing targets using I-specific cleavage and exon array analysis. Mol Cell Probes 7, 38&ndash;45. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/22960667" target="_blank">22960667</a></p> <p>Wirth MJ and Wahle P (2003). Biolistic transfection of organotypic cultures of rat visual cortex using a handheld device. J Neurosci Methods 125, 45&ndash;54. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/12763229" target="_blank">12763229</a></p> <p>Zhang G and Selzer ME (2001). In vivo transfection of lamprey brain neurons by gene gun delivery of DNA. Exp Neurol 167, 304&ndash;311. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/11161618" target="_blank">11161618</a></p> <div class="top"><a href="#helptop">Back to Top</a></div> Selection guides <table id="carttablealigned" class="literature_table" style="height: auto; width: 583px;" border="0" cellspacing="0" cellpadding="0"> <tbody> <tr> <td width="100">6176</td> <td width="350">Electroporation Systems Overview</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6176.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6177</td> <td width="350">Biolistic Particle Delivery Systems</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6177.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6178</td> <td width="350">Recommended Biolistic System by Cell Types</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6178.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6179</td> <td width="350">Lipid Transfection Reagents Selection Guide</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6179.pdf" target="_blank"><span>Click to download</span></a></td> </tr> </tbody> </table> <div class="videowrap vwrap_last"> <div class="videoImg"><a title="Gene Pulser Xcell Electroporation System: Components, Application, and Troubleshooting" onclick="window.open('/webroot/web/movies/lsr/support/Gene_Pulser_Xcell.htm', 'StatusBar', 'toolbar=no, resizable=no, scrollbars=no, width=810, height=575');" href="javascript:void(0);"><img style="border:none;" src="https://www.bio-rad.com/webroot/web/images/lsr/support/tutorials/global/xcell_tn.png" alt="" /></a></div> <div class="videoDesc"><a title="Gene Pulser Xcell Electroporation System: Components, Application, and Troubleshooting" onclick="window.open('/webroot/web/movies/lsr/support/Gene_Pulser_Xcell.htm', 'StatusBar', 'toolbar=no, resizable=no, scrollbars=no, width=810, height=575');" href="javascript:void(0);">Gene Pulser Xcell&trade; Electroporation System: Components, Application, and Troubleshooting</a><br /> This tutorial highlights the main components and features of the Gene Pulser Xcell system. It provides information about system installation and the setup of electroporation experiments, including important troubleshooting tips and answers to frequently asked questions. Ordering information for system components and accessories is also provided.</div> <div class="clear">&nbsp;</div> </div> 5448 5542 5582 5399 5445 5598 5634 1908 5555 5554 5924 1349 1365 1345 1355 1365 1345 1349 5542 5858 5641 5603 5622 5686 5687 5720 5704 5733 5774 5778 5823 5842 5684 0108 5860 5904 2497 5443 5447 5446 2051 2015 2007 2087 2658 2453 1688 1689 2433 2552 2453 2531 2410 2726 1689 2768 Life Science Research/Products/Transfection/Electroporation/Gene Pulser Xcell Electroporation Systems ->MT::b1a35eb3-d55c-47b3-aaf3-95e4d1d85848##Life Science Research/Products/Transfection/Electroporation/Gene Pulser MXcell Electroporation System ->MT::fae0f825-da45-4b02-aa33-73781dda6171##Life Science Research/Products/Transfection/Electroporation/MicroPulser Electroporator ->MT::83527990-34fb-4b33-b955-ca53b57bf8b9##Life Science Research/Products/Transfection/Biolistic Particle Delivery Systems/Helios Gene Gun Systems ->MT::42e9d6be-369a-49f8-8fbb-281a0fea6df8##Life Science Research/Products/Transfection/Biolistic Particle Delivery Systems/PDS-1000 | He and Hepta Systems ->MT::1730e08d-f43a-46ea-b7f3-7b35c04c36eb## Life Science Research/Solutions/Technologies/qPCR|Real-Time PCR ->MTS::LUSO4W8UU##Life Science Research/Solutions/Technologies/Cell Counting Methods ->MTS::LUSOLB470##Life Science Research/Solutions/Technologies/Imaging and Analysis ->MTS::LUSQC6MNI##Life Science Research/Solutions/Technologies/Transfection ->MTS::LUSONCB9O## Eddie C Instrument-Based Transfection Methods 12/28/11 04:27 PM 12/28/21 04:37 PM AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA en LSR /LSR/Technologies/Transfection N 0
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