iScript™ Advanced cDNA Synthesis Kit for RT-qPCR

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iScript™ Advanced cDNA Synthesis Kit

Overview

iScript™ Advanced cDNA Synthesis Kit for RT-qPCR Workflow
iScript™ Advanced cDNA Synthesis Kit for RT-qPCR Workflow
Extended Dynamic Range and Higher Capacity
Extended dymanic range and higher capacity

 

Total RNA (7.5 µg–1 pg) from HeLa cells was reverse transcribed using the iScript™ advanced cDNA synthesis kit for RT-qPCR in a 20 µl reaction. A tenfold dilution of generated cDNA was used as template to amplify α-tubulin in a 10 µl qPCR reaction with iQ™ SYBR® Green supermix on a CFX384™ real-time PCR detection system. Efficiency = 90.7%, R2 = 0.999, slope = –3.57. Cq, quantification cycle; RFU, relative fluorescence units.

Accurate mRNA Representation
Accurate mRNA Representation

 

Total RNA (7.5, 5, 2.5, and 1 µg) from human spleen was reverse transcribed using the iScript™ advanced cDNA synthesis kit for RT-qPCR. The generated cDNA samples were diluted to the equivalent of 50 ng/µl and used as template to amplify a low-expressed gene (IL-2) in a 10 µl qPCR reaction with SsoFast™ probes supermix on a CFX384™ real-time PCR detection system (FAM probe). There were no significant differences (<0.5 Cq) between diluted samples, which demonstrates accurate representation of mRNA transcripts for high and low input RNA. RFU, relative fluorescence units.

Efficient Amplification of High GC Targets
Efficient Amplification of High GC Targets

Six different input RNA amounts (7.5, 5, 2, 1, 0.1, and 0.01 µg) from HeLa cells were reverse transcribed using the iScript™ advanced cDNA synthesis kit for RT-qPCR in a 20 µl reaction. The generated cDNA was diluted 20-fold for qPCR. A 110 bp CBP amplicon (70% GC) was amplified in a 10 µl qPCR reaction with SsoFast™ EvaGreen® supermix on a CFX96™ real-time PCR detection system. CBP efficiency = 99%, R2 = 0.998, slope = –3.33. RFU, relative fluorescence units.

Superior 5' and 3' Coverage
Superior 5' and 3' Coverage

Total RNA (7.5, 2, 1, and 0.1 µg) from HeLa cells was reverse transcribed using the iScript™ advanced cDNA synthesis kit for RT-qPCR in a 20 µl reaction. One-tenth of the generated cDNA was used as template to amplify the APC gene in a 10 µl qPCR reaction with iQ™ SYBR® Green supermix on a CFX384™ system. Primer pairs were designed at the 5' end () and 3' end () for 91 and 69 bp amplicons. There was no significant difference between the two primer pairs, which demonstrates superior coverage of the 5' and 3' regions of target mRNA. RFU, relative fluorescence units.

Which iScript™ Kit is right for me?
 

Most
Convenient

Integrated
DNA
Removal

Maximum
Input Use

Flexible Priming,
High-Fidelity

Value
Solution

Product iScript Reverse Transcription Supermix for RT-qPCR iScript gDNA
Clear cDNA
Synthesis Kit
iScript Advanced cDNA Synthesis Kit for RT-qPCR iScript Select cDNA
Synthesis Kit
iScript cDNA
Synthesis Kit
Applications
  • Gene expression
  • Gene expression
  • Gene expression
  • Cloning
  • Sequencing and NGS
  • Gene expression
  • Gene expression
Total Input RNA 1 µg–1 pg 1 µg–1 pg 7.5 µg–100 fg 1 µg–1 pg 1 µg–100 fg
Format 1 tube 3 tubes 2 tubes 5 tubes 2 tubes
Kit Contents
  • 5x iScript RT Supermix
  • No-RT control supermix
  • 5x iScript RT Supermix
  • No-RT control supermix
  • DNase
  • DNase buffer
  • iScript Reverse Transcriptase
  • 5x iScript Advanced Reaction Mix
  • iScript Reverse Transcriptase
  • 5x iScript Reaction Mix
  • 3 priming options
  • iScript Reverse Transcriptase
  • 5x iScript Reaction Mix
Time to Produce cDNA 26 min 36 min 21 min 15–65 min 26 min
RNase H+ Activity High High High High High
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The iScript Advanced cDNA Synthesis Kit for RT-qPCR is an enhanced formulation that delivers increased data throughput from a single 20 µl reverse transcription (RT) reaction for real-time qPCR. This two-tube kit enables superior RT capacity of input RNA up to 7.5 µg.

Key Features and Benefits

  • Increase qPCR data throughput and cost effectiveness from a single reaction — cDNA synthesized from higher input RNA amounts can be used for analyzing large numbers of target genes
  • Detect low-level target genes — uncompromised sensitivity even with less input RNA for when RNA sample is limited
  • Reduce inter-assay variability and improve reproducibility — eliminate the need for multiple cDNA synthesis reactions
  • Allow true representation of target mRNA levels — potent blend of RNase inhibitor prevents RNA degradation during reaction setup and RT
  • Increase primer design flexibility and prevent 5′ and 3′ bias — optimum blend of oligo(dT) and random primers for superior RNA coverage
  • Set up reactions with ease and obtain cDNA in less time — 2-tube kit generates cDNA in 21 minutes
Storage at –20°C Guaranteed for 12 months at –20°C in a constant temperature freezer (multiple freeze thaws not recommended)
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iScript™ Advanced cDNA Synthesis Kit, 25 x 20 µl rxns

1725037
25 x 20 µl reactions, includes 100 µl iScript Advanced 5x Reaction Mix, 25 µl iScript Advanced Reverse Transcriptase

1725037 /templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5037_1401382449.xml 1725037 05/29/14 09:45 AM en_US true internet/product_SKU 2018-02-27 06:49:41 /default/main/bio-rad/STAGING 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 templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5037_1401382449.xml /default/main/bio-rad/WORKAREA/default jdl0szao 2017-11-14 07:07:36 true jdl0szao templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5037_1401382449.xml LSR N /default/main/bio-rad en 05/29/24 09:53 AM 1519353027361 2018-02-27 06:49:41 /default/main/bio-rad/WORKAREA/default/templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5037_1401382449.xml 1725037-iscript-advanced-cdna-synthesis-kit-25-x-20-ul-rxns 1 1725037 iScriptAdvancedcDNASynthesisKitforRT-qPCR iScript™ Advanced cDNA Synthesis Kit, 25 x 20 µl rxns 25 x 20 µl reactions, includes 100 µl iScript Advanced 5x Reaction Mix, 25 µl iScript Advanced Reverse Transcriptase 25 x 20 µl reactions, includes 100 µl iScript Advanced 5x Reaction Mix, 25 µl iScript Advanced Reverse Transcriptase /webroot/web/images/lsr/products/amplification_pcr/sku_thumb/global/170-8842_tn.jpg /webroot/web/images/lsr/products/amplification_pcr/sku_view/global/170-8842_view.jpg /webroot/web/images/lsr/products/amplification_pcr/sku_feature/global/170-8842_feature.jpg iScript Advanced cDNA Synthesis Kit for RT-qPCR preview Dave Lubertozzi iScript Advanced cDNA Synthesis Kit 1725037, iscript, advanced, cdna, synthesis, rt-qpcr, 25x20, reactions, reverse, transcription, first, strand, complimentary, gene, expression, real, time, qpcr, detection, real-time, pcr 05/29/14 09:45 AM 05/29/24 09:53 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 en LSR /LSR N 0 <p>Use the iScript Advanced cDNA Synthesis Kit for RT-qPCR for increased data throughput from a single 20 &micro;l reverse transcription (RT) reaction for real-time qPCR. Two-tube kit allows up to 7.5 &micro;g input RNA.</p> <ul> <li>Ability to use higher input RNA amounts to analyze a large number of target genes</li> <li>Detect low-level target genes &mdash; use with less input RNA, or when RNA sample is limited</li> <li>Reduce inter-assay variability and improve reproducibility &mdash; eliminate the need for multiple cDNA synthesis reactions</li> <li>Prevent 5&prime; and 3&prime; bias &mdash; optimum blend of oligo(dT) and random primers</li> <li>25 &micro;l (enough for 25 x 20 &micro;l reactions)</li> <li>2-tube kit generates cDNA in 21 min</li> </ul> <h2 class="descHeader">Packaging Options</h2> <div class="clear" style="height: 1px;">&nbsp;</div> <table class="pd_table pd_gridlines" border="0"> <tbody> <tr> <td align="left"><strong>Kit Size</strong></td> <td align="left"><strong>Catalog Number</strong></td> </tr> <tr class="pd_colorbackground"> <td align="left" valign="top">25 x 20 &micro;l reactions</td> <td align="left" valign="top">172-5037</td> </tr> <tr> <td align="left" valign="top">100 x 20 &micro;l reactions</td> <td align="left" valign="top"><a style="font-size: 11px;" href="/en-us/sku/172-5038-iscript-advanced-cdna-synthesis-kit-for-rt-qpcr">172-5038</a></td> </tr> </tbody> </table> <br /> <h2 class="descHeader">Related Products</h2> <ul> <li><a href="/evportal/destination/commerce/product_detail?catID=M87EVMKG4"><strong><span style="text-decoration: underline;">iScript&trade; Reverse Transcription Supermix for RT-qPCR</span></strong></a>: 1-tube format for total input 1 &micro;g&ndash;1 pg RNA, plus liquid at &ndash;20&deg;C, eliminating freeze-thaw cycles</li> <li><a href="/evportal/destination/commerce/product_detail?catID=M87EZXC4S"><strong><span style="text-decoration: underline;">iScript&trade; Select cDNA Synthesis Kit</span></strong></a>: synthesize long cDNAs up to 8 kb; oligo(dT), random primer, and gene-specific primer enhancer provided in separate tubes</li> <li><a href="/evportal/destination/commerce/product_detail?catID=M87EWZESH"><strong><span style="text-decoration: underline;">iScript&trade; cDNA Synthesis Kit</span></strong></a>: 2-tube format allows for up to 15 &micro;l of RNA sample in a 20 &micro;l reaction, in a 40 min protocol</li> </ul> <h2 class="descHeader">More Information</h2> <p>Learn about the <a href="/en-us/product/reverse-transcription-reagents/iscript-advanced-cdna-synthesis-kit-for-rt-qpcr?tab=Description">key features and specifications of the iScript&trade; Advanced cDNA Synthesis Kit for RT-qPCR.</a></p> /webroot/web/html/lsr/products/SKUs/autopopulated 172-5037.html /webroot/web/html/lsr/products/SKUs/autopopulated/172-5037.html 1 0 false false false /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725037&CartQuote=QT&quantity= false true false $196.00 false null /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725037&AddToDefaultWishList=true&quantity= true false true true false true true true /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725037&quantity= 1725037
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iScript™ Advanced cDNA Synthesis Kit, 100 x 20 µl rxns

1725038
100 x 20 µl reactions, includes 400 µl 5x iScript Advanced Reaction Mix, 100 µl iScript Advanced Reverse Transcriptase

1725038 /templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5038_1403297461.xml 1725038 06/20/14 01:43 PM en_US true internet/product_SKU 2018-02-27 06:49:38 /default/main/bio-rad/STAGING 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 templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5038_1403297461.xml /default/main/bio-rad/WORKAREA/default jcihuc20 2017-11-14 07:07:36 true jcihuc20 templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5038_1403297461.xml LSR N /default/main/bio-rad en 06/20/24 01:45 PM 1519353027407 2018-02-27 06:49:38 /default/main/bio-rad/WORKAREA/default/templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5038_1403297461.xml 1725038-iscript-advanced-cdna-synthesis-kit-100-x-20-ul-rxns 1 1725038 iScriptAdvancedcDNASynthesisKitforRT-qPCR iScript™ Advanced cDNA Synthesis Kit, 100 x 20 µl rxns 100 x 20 µl reactions, includes 400 µl 5x iScript Advanced Reaction Mix, 100 µl iScript Advanced Reverse Transcriptase 100 x 20 µl reactions, includes 400 µl 5x iScript Advanced Reaction Mix, 100 µl iScript Advanced Reverse Transcriptase /webroot/web/images/lsr/products/amplification_pcr/sku_thumb/global/170-8842_tn.jpg /webroot/web/images/lsr/products/amplification_pcr/sku_view/global/170-8842_view.jpg /webroot/web/images/lsr/products/amplification_pcr/sku_feature/global/170-8842_feature.jpg iScript Advanced cDNA Synthesis Kit for RT-qPCR preview Dave Lubertozzi 1725038, iscript, advanced, cdna, synthesis, rt-qpcr, 100x20, reactions, reverse, transcription, first, strand, complimentary, gene, expression, real-time, real, time, pcr, qpcr, detection 06/20/14 01:43 PM 06/20/24 01:45 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/Amplification---PCR/PCR-Reagents N 0 <p>Use the iScript Advanced cDNA Synthesis Kit for RT-qPCR for increased data throughput from a single 20 &micro;l reverse transcription (RT) reaction for real-time qPCR. Two-tube kit allows up to 7.5 &micro;g input RNA.</p> <ul> <li>Ability to use higher input RNA amounts to analyze a large number of target genes</li> <li>Detect low-level target genes &mdash; use with less input RNA, or when RNA sample is limited</li> <li>Reduce inter-assay variability and improve reproducibility &mdash; eliminate the need for multiple cDNA synthesis reactions</li> <li>Prevent 5&prime; and 3&prime; bias &mdash; optimum blend of oligo(dT) and random primers</li> <li>25 &micro;l (enough for 25 x 20 &micro;l reactions)</li> <li>2-tube kit generates cDNA in 21 min</li> </ul> <h2 class="descHeader">Packaging Options</h2> <div class="clear" style="height: 1px;">&nbsp;</div> <table class="pd_table pd_gridlines" border="0"> <tbody> <tr> <td align="left"><strong>Kit Size</strong></td> <td align="left"><strong>Catalog Number</strong></td> </tr> <tr class="pd_colorbackground"> <td align="left" valign="top">25 x 20 &micro;l reactions</td> <td align="left" valign="top"><a href="/en-us/sku/172-5037-iscript-advanced-cdna-synthesis-kit-for-rt-qpcr">172-5037</a></td> </tr> <tr> <td align="left" valign="top">100 x 20 &micro;l reactions</td> <td align="left" valign="top"><span style="font-size: 11px;">172-5038</span></td> </tr> </tbody> </table> <br /> <h2 class="descHeader">Related Products</h2> <ul> <li><a href="/evportal/destination/commerce/product_detail?catID=M87EVMKG4"><strong><span style="text-decoration: underline;">iScript&trade; Reverse Transcription Supermix for RT-qPCR</span></strong></a>: 1-tube format for total input 1 &micro;g&ndash;1 pg RNA, plus liquid at &ndash;20&deg;C, eliminating freeze-thaw cycles</li> <li><a href="/evportal/destination/commerce/product_detail?catID=M87EZXC4S"><strong><span style="text-decoration: underline;">iScript&trade; Select cDNA Synthesis Kit</span></strong></a>: synthesize long cDNAs up to 8 kb; oligo(dT), random primer, and gene-specific primer enhancer provided in separate tubes</li> <li><a href="/evportal/destination/commerce/product_detail?catID=M87EWZESH"><strong><span style="text-decoration: underline;">iScript&trade; cDNA Synthesis Kit</span></strong></a>: 2-tube format allows for up to 15 &micro;l of RNA sample in a 20 &micro;l reaction, in a 40 min protocol</li> </ul> <h2 class="descHeader">More Information</h2> <p>Learn about the <a href="/en-us/product/reverse-transcription-reagents/iscript-advanced-cdna-synthesis-kit-for-rt-qpcr?tab=Description">key features and specifications of the iScript&trade; Advanced cDNA Synthesis Kit for RT-qPCR.</a></p> /webroot/web/html/lsr/products/SKUs/autopopulated 172-5038-1.html /webroot/web/html/lsr/products/SKUs/autopopulated/172-5038-1.html 1 0 false false false /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725038&CartQuote=QT&quantity= false true false $618.00 false null /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725038&AddToDefaultWishList=true&quantity= true false true true false true true true /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725038&quantity= 1725038
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Number Description Options
10000070791 iScript™ Advanced cDNA Synthesis Kit for RT-qPCR Product Insert, Ver B Click to download
6125 iScript™ Advanced cDNA Synthesis Kit for RT-qPCR Product Information Sheet, Rev A Click to download [ Add to Cart (Free) ]
6746 iScript™ Reverse Transcription Kits Flier, Ver B Click to download
6252 Reagent Comparison Guide for Real-Time PCR Brochure, Rev C Click to download [ Add to Cart (Free) ]
6090 Amplification Reagents and Plastics Brochure Click to download [ Add to Cart (Free) ]
6763 Gene Expression Real-Time PCR Workflow Flier, Ver A Click to download [ Add to Cart (Free) ]
6777 Protocol for Single Cell Gene Expression Profiling by Multiplex RT-qPCR, Ver A Click to download
6816 Protocol for Multiplex RT-qPCR Analysis Using PrimePCR™ Assays, Ver A Click to download
6833 High Throughput Single Cell Gene Expression Profiling by Multiplex qPCR, Ver A Click to download

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