Digital PCR Library Quantification Kits

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Digital PCR Library Quantification Kits

Overview

QX200™ Droplet Digital™ PCR System Brochure

Droplet Digital PCR (ddPCR™) is Bio-Rad's unique digital PCR technology.

  • Unrivaled precision provides absolute quantification of target DNA or RNA molecules without the use of standard curves
  • Addresses the lack of scalable and practical technologies for digital PCR implementation

The QX200 ddPCR system makes this powerful technology accessible to you for unveiling previously unattainable
research levels.

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QX200™ Droplet Digital™ (ddPCR™) System Data
Detection of BRAF V600E Mutation

High sensitivity of the QX200 system allows quantification of BRAF V600E mutation in the presence of wild-type DNA using PrimePCR ddPCR assays. A, 2-D fluorescence amplitude plot generated by QuantaSoft™ software shows triplicate wells of a mixed mutant:wild-type sample. The black cluster on the plot represents the negative droplets, the green cluster represents the droplets that are positive for wild-type DNA only, the blue cluster represents the droplets that are positive for mutant DNA only, and the orange cluster represents the droplets that are positive for both mutant and wild-type DNA. B, 2-D fluorescence amplitude plot shows three replicates of a wild-type-only sample. C, fractional abundance plot shows the percentage frequency (orange markers) of the mutant DNA in a wild-type DNA background. The blue markers indicate the concentration of mutant DNA (copies/µl) and the green markers indicate the concentration of wild-type DNA (copies/µl) in each of three replicate samples. All error bars generated by QuantaSoft software represent the 95% confidence interval.



Detection of Herpes Simplex Virus and β2 Microglobulin

ddPCR enables precise and reproducible detection of herpes simplex virus 1 (HSV-1) and ß2 microglobulin (B2M) targets in a duplex assay. A, 2-D fluorescence amplitude plot shows three merged replicate samples of 10,000 copies/well of HSV-1 duplexed with 20,000 copies/well of Β2M. The black cluster on the plot represents the negative droplets, the green cluster represents the droplets that are positive for HSV-1 only, the blue cluster represents the droplets that are positive for B2M only, and the orange cluster represents the droplets that are positive for both HSV-1 and B2M targets. B, concentration plot shows merged triplicate wells across an HSV-1 sample dilution series from 0 to 40,000 copies in a constant background of 20,000 copies (66 ng human genomic DNA) of B2M. The blue markers indicate HSV-1 copies/µl and the green markers indicate B2M copies/µl. All error bars generated by QuantaSoft software represent the 95% confidence interval.



Detection of Copy Number for
Myelocytomatosis Oncogene and β-Globin

The QX200 system provides superior resolution with copy number (CN) calls from CN1 to CN13. A, 2-D fluorescence amplitude plot shows duplicate wells of a CN6 sample duplexed with myelocytomatosis oncogene (MYC) and β-globin (HBB) assays. The black cluster on the plot represents the negative droplets, the green cluster represents the droplets that are positive for MYC only, the blue cluster represents the droplets that are positive for HBB only, and the orange cluster represents the droplets that are positive for both MYC and HBB targets. B, copy number plot shows samples ranging from CN1 to CN13 with precise duplicate well values. All error bars generated by QuantaSoft software represent the 95% confidence interval. NTC, no template control.



Validation for Next-Generation Sequencing

ddPCR provides nearly 1,000-fold greater sensitivity and an amplification bias–free approach compared to RNA-Seq for validation. A, concentration plot shows a twofold dilution of cDNA generated using Bio-Rad’s iScript™ advanced cDNA synthesis kit for RT-qPCR. Two independent measurement sets were made: one at a high concentration range and one at a lower range, with four points overlapping. B, table shows comparative results between ddPCR and MiSeq. ddPCR detected thousands of copies per well with low-abundance transcripts (TBP and GUSB genes) using an input of 100 ng total RNA, which demonstrates enhanced sensitivity. The MiSeq sequencer detected only single-digit reads per kilobase per million reads (RPKM) for these transcripts. ddPCR, Droplet Digital PCR.



Gene Expression Analysis of Stem Cell Markers

High sensitivity and precision of the QX200 system allow reliable detection of small fold changes with rare transcripts. The TGFβ signaling pathway plays a role in ovarian stem cell regulation, and the addition of TGFβ will increase the number of stem cells in the population, increase the expression of putative stem cell markers, and decrease terminal differentiation markers like E-cadherin (ECAD). Data show expression levels of cKIT, CD133, OCT3/OCT4, and ECAD genes across ten samples with and without TGFβ treatment, normalized to PPIA in this study. Reproducible detection of cDNA copies as low as 0.5–1 target/µl in a 20 µl reaction was achieved. S, sample.


Digital PCR using QX200™ and AutoDG™ ddPCR Systems
Success Stories: Articles and Videos

Success Stories: Articles

Genomic Variation Analysis: Single Nucleotide Polymorphism: Validation of Allele-Specific Expression Predicted by RNA-Seq in Human Brain Specimens
In the context of human brain development, the QX100™ Droplet Digital™ PCR System yields unambiguous results for genotype identification of low-expression genes and provides absolute quantification of allele-specific expression (ASE) without the use of a standard.
Download the article.

Genomic Variation Analysis: Copy Number Variation: DUF1220 Copy Number Variation across Healthy and Diseased Populations
Studying copy number variation (CNV) with the QX100 Droplet Digital PCR System allows the robust identification of genetic factors such as DUF1220 in neurogenetic diseases between individuals with greater accuracy and precision, while effectively validating array CGH data.
Download the article.

Genomic Variation Analysis: Single Nucleotide Polymorphism: Detection of Single Nucleotide Variation Frequencies by Droplet Digital™ PCR
The QX100 Droplet Digital PCR System validated DNA sequencing results of induced pluripotent stem cells (iPSCs). In addition to confirming the high frequency of single nucleotide variations (SNVs), a de novo type of SNV was identified.
Download the article.

Cancer Mutation Analysis: How Droplet Digital PCR Has Revolutionized Noninvasive Plasma-Based Detection of Mutations
Performing Droplet Digital PCR with the QX100 Instrument effectively detected and accurately quantified the EGFR T790M marker in targeted therapies with limited cancer tumor samples.
Download the article.

Viral Quantification: Adeno-Associated Virus Vector Genome Titer Assay
Determining the adeno-associated virus (AAV) vector genome (vg) titer assay is critical in gene therapy applications. The improved sensitivity observed with Droplet Digital PCR (ddPCR™) technology makes it highly suitable for titration of clinical-grade AAV vector preparations.
Download the article.

Gene Expression: Digital Quantification of Potential Therapeutic Target RNAs
Measuring the exact number of long noncoding RNAs (lncRNAs) in subcellular compartments with Droplet Digital PCR (ddPCR) provides insights into their role and mechanism of action.
Download the article.

Third Generation PCR: A Novel Way to Accurately Measure Copy Number Variations in Human Genome
The McCarroll Lab uses Droplet Digital technology to study the distribution and molecular properties of the genome and its influence on gene expression, as well as the genetics underlying schizophrenia and bipolar disorder.
Download the article.

Success Stories: Videos

Droplet Digital™ PCR opens new perspectives in HIV research. Sangamo BioSciences, Inc. is developing HIV therapeutics toward a functional cure using the company’s proprietary zinc finger DNA-binding protein technology.
Download the article.


Professor Alec Morley, a pioneer in digital PCR, and his colleagues in Adelaide have developed tests for detecting acute lymphoblastic leukemia (ALL) and chronic myeloid leukemia (CML) using the unrivaled sensitivity of the QX200 Droplet Digital PCR System.



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Precise quantification of next-generation sequencing (NGS) libraries is critical for the efficient use of NGS platforms. Bio-Rad's digital PCR platform and library quantification kits can be easily incorporated into the NGS library preparation workflow to precisely quantify and balance sequencing libraries on the Illumina or Ion Torrent Platforms.

The ddPCR library quantification assay generates data plots that are rich with qualitative library information, a feature not available with other current methodologies.

When paired with the QX200 Droplet Digital PCR System, these kits allow accurate quantification and provide a qualitative measure of the DNA library prior to sequencing on Illumina (186-3040).

Features of Digital PCR Library Quantification Kits

  • Contain all the necessary components to create droplets and quantify NGS libraries
  • Provide information about library quality, such as adapter dimers, and indicate library insert size
  • Provide more efficient and consistent loading of libraries for sequencing runs
  • Enable balancing of pooled library samples
  • Optimize the use of consumables, labor, and instrument time for the highest productivity with NGS

Related Products

Bio-Rad offers additional digital PCR supermixes including:

Additional Equipment

These digital PCR assays require the use of the following instruments and accessories:

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(multiple freeze/thaws not recommended)
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ddPCR™ Library Quantification Kit for Illumina TruSeq

1863040
200 x 20 µl reactions, includes ddPCR Library Quantification Assay and ddPCR Supermix for Probes (No dUTP), for quantification of Illumina TruSeq libraries using the QX200™/QX100™ ddPCR™ Systems

1863040 /templatedata/internet/product_SKU/data/LSR/Amplification---PCR/Droplet-Digital-PCR-System/186-3040_1365010259.xml 1863040 /default/main/bio-rad/WORKAREA/default 04/03/13 09:29 AM en_US jc7v8odn 2017-11-14 07:07:29 true true internet/product_SKU jc7v8odn templatedata/internet/product_SKU/data/LSR/Amplification---PCR/Droplet-Digital-PCR-System/186-3040_1365010259.xml LSR 2019-05-05 00:54:33 /default/main/bio-rad/STAGING N /default/main/bio-rad en 04/03/23 10:20 AM 1555973500764 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,VN 2019-05-05 00:54:33 /default/main/bio-rad/WORKAREA/default/templatedata/internet/product_SKU/data/LSR/Amplification---PCR/Droplet-Digital-PCR-System/186-3040_1365010259.xml templatedata/internet/product_SKU/data/LSR/Amplification---PCR/Droplet-Digital-PCR-System/186-3040_1365010259.xml 1863040-ddpcr-library-quantification-kit-for-illumina-truseq 1863040 ddPCR? Library Quantification Kit for Illumina TruSeq ddPCR&#153; Library Quantification Kit for Illumina TruSeq 200 x 20 &micro;l reactions, includes ddPCR Library Quantification Assay and ddPCR Supermix for Probes (No dUTP), for quantification of Illumina TruSeq libraries using the QX200&trade;/QX100&trade; ddPCR&trade; Systems 200 x 20 &micro;l reactions, includes ddPCR Library Quantification Assay and ddPCR Supermix for Probes (No dUTP), for quantification of Illumina TruSeq libraries using the QX200&trade;/QX100&trade; ddPCR&trade; Systems /webroot/web/images/lsr/products/amplification_pcr/sku_thumb/global/ddpcr-library-quantification-kit-for-ilumina-truseq-186-3040-thumb.jpg /webroot/web/images/lsr/products/amplification_pcr/sku_view/global/ddpcr-library-quantification-kit-for-ilumina-truseq-186-3040-view.jpg ,VN Eddie C ddPCR&#153; Library Quantification Kit for Illumina TruSeq 200 x 20 &micro;l reactions, includes ddPCR Library Quantification Assay and ddPCR Supermix for Probes (No dUTP), for quantification of Illumina TruSeq libraries using the QX200&trade;/QX100&trade; ddPCR&trade; Systems 1863040, ddpcr, library, quantification, illumina, truseq, quantitation, amplification, assay, supermix, probes, libraries, qx200, qx100, droplet, digital, pcr, instrument 04/03/13 09:29 AM 04/03/23 10:20 AM 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,VN en LSR /LSR/Amplification---PCR/Droplet-Digital-PCR-System N 0 <p>Use the <strong>ddPCR Library Quantification Kit</strong> to quantify the sequencing libraries generated by the Illumina<br /> TruSeq Kit using the <a href="/evportal/destination/commerce/product_detail?catID=MPOQQE4VY">QX200&trade; Droplet Digital&trade; PCR System</a>.</p> <h2 class="descHeader">Kit Components</h2> <ul> <li>ddPCR Quantification Assay &mdash; 1 x 200 &micro;l</li> <li>ddPCR Supermix for Probes (no dUTP) &mdash; 2 x 1 ml</li> </ul> <h2 class="descHeader">Kit Features and Benefits</h2> <ul> <li><strong>Superior performance</strong> &mdash; generates more qualitative, information-rich data with no PCR bias</li> <li><strong>Simple workflow</strong> &mdash; easily incorporated into the current NGS library construction and QC workflow </li> <li><strong>Efficient utilization of sequencing runs</strong> &mdash; provides fluorescence amplitude data that correlate with library quality</li> </ul> <h2 class="descHeader">Related Products</h2> <p>Bio-Rad also offers the ddPCR Library Quantification Kit for the Ion Torrent Platform (<a href="/evportal/destination/commerce/sku_detail?cmd=catProductDetail&amp;vertical=LSR&amp;productID=186-3041">186-3041</a>).</p> <p>In addition, <a href="/evportal/destination/product?catID=N3D44215">digital PCR supermixes</a> are available for completing your digital PCR workflow:</p> <ul> <li><a href="/evportal/destination/commerce/product_detail?catID=N3D45ZE8Z">ddPCR&trade; Supermix for Probes (no dUTP)</a></li> <li><a href="/evportal/destination/commerce/product_detail?catID=N3D47S4VY">ddPCR&trade; Supermix for Probes</a></li> <li><a href="/evportal/destination/commerce/product_detail?catID=N3D48FKG4">QX200&trade; ddPCR&trade; EvaGreen Supermix</a></li> </ul> <h2 class="descHeader">More Information</h2> <p>Learn more about the features of our <a href="/evportal/destination/commerce/product_detail?catID=N6EOPN15">digital PCR library quantification kits</a>.</p> /webroot/web/html/lsr/products/amplification_pcr/sku_view/description/global ddpcr-ibrary-quantification-kit-for-illumina-truseq-186-3040-desc.html /webroot/web/html/lsr/products/amplification_pcr/sku_view/description/global/ddpcr-ibrary-quantification-kit-for-illumina-truseq-186-3040-desc.html Storage at –20°C 2199 Guaranteed up to 18 months in a constant temperature freezer <br> (multiple freeze/thaws not recommended) 1 0 false false false /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1863040&CartQuote=QT&quantity= 1357 false true false $985.00 false null /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1863040&AddToDefaultWishList=true&quantity= true false true 41106300 true false true true true /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1863040&quantity= 1863040
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Number Description Options
6402 ddPCR Library Quantification Kit for Illumina TruSeq Flier, Rev B Click to download
10031986 ddPCR Library Quantification Kit for Illumina TruSeq Product Insert, Rev C Click to download
6343 QX100 Droplet Digital PCR System for Accurate Quantification of Illumina TruSeq Libraries, Rev B Click to download
6490 ddPCR Library Quantification Kit for Ion Torrent Flier, Rev A Click to download
10033630 ddPCR Library Quantification Kit for Ion Torrent Product Insert, Rev B Click to download
6495 Bio-Rad’s ddPCR™ Library Quantification Kit for Ion Torrent Enables Accurate Library Quantification and Excellent Balancing of Pooled Libraries, Rev A Click to download
6407 Droplet Digital PCR Applications Guide Click to download
6423 Droplet Digital PCR: Ultra-Sensitive Validation Technology for RNA-Seq Application Note, Rev A Click to download [ Add to Cart (Free) ]
10023997 Instruction Manual, PX1 PCR Plate Sealer, Rev A Click to download

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