iScript™ Advanced cDNA Synthesis Kit for RT-qPCR

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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?


iScript Kit Selector

iScript™ Reverse Transcription Supermix for RT-qPCR iScript™ Advanced cDNA Synthesis Kit for RT-qPCR iScript™ cDNA Synthesis Kit iScript™ Select cDNA Synthesis Kit iScript™ One-Step RT-PCR Kit With SYBR® Green iScript™ One-Step RT-PCR Kit for Probes

Coming Soon — New Packaging Sizes
iScript™ Advanced cDNA synthesis kit packaging sizes will soon be updated to better suit your needs; see the table below for details.

 
Current Kit Size Catalog Number New Kit Size Catalog Number
50 x 20 µl reactions 170-8842 25 x 20 µl reactions (coming soon)
250 x 20 µl reactions 170-8843 100 x 20 µl reactions (coming soon)

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 35 min
Storage at –20°C Guaranteed for 12 months at –20°C in a constant temperature freezer (multiple freeze thaws not recommended)
iScript Advanced cDNA Synthesis Kit for RT-qPCR
iScript™ Advanced cDNA Synthesis Kit for RT-qPCR

170-8842
50 x 20 µl reactions, includes 200 µl 5x reaction mix, 50 µl iScript advanced reverse transcriptase
Product size soon to be changed

List Price:   $369.00
Quantity:   Add to cart
Add to Hot List   Add to Quote
iScript™ Advanced cDNA Synthesis Kit for RT-qPCR

170-8843
250 x 20 µl reactions, includes 1,000 µl 5x iScript advanced reaction mix, 250 µl iScript advanced reverse transcriptase
Product size soon to be changed

List Price:   $1,487.00
Quantity:   Add to cart
Add to Hot List   Add to Quote
Number Description Options
6252 Reagent Comparison Guide for Real-Time PCR Brochure, Rev C Click to download
6090 Amplification Reagents and Plastics Brochure, Rev D Click to download [ Add to Cart (Free) ]
6125 iScript Advanced cDNA Synthesis Kit for RT-qPCR Product Information Sheet, Rev A Click to download [ Add to Cart (Free) ]
10022089 Instruction Manual, iScript Advanced cDNA Synthesis Kit for RT-qPCR, Rev A Click to download