iScript™ Select cDNA Synthesis Kit

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Overview

Broad Dynamic Range

The iScript™ Select cDNA synthesis kit performs reliably over six orders of magnitude using any first-strand priming approach. Human total RNA from 1 pg to 1 µg was reverse transcribed using the iScript Select cDNA synthesis kit and the first-strand primers indicated above. The resulting cDNA was amplified in triplicate using iQ™ SYBR® Green supermix with β-actin primers and detected using the iCycler iQ® real-time detection system, RFU, relative fluorescence units.

Long cDNA Synthesis

The iScript Select cDNA synthesis kit facilitates synthesis of cDNA longer than 6 kb. First-strand cDNA was produced from 1 µg total RNA using the iScript Select cDNA synthesis kit and the provided oligo(dT) primer mix. A 2 µl aliquot of the 20 µl cDNA synthesis reaction was subjected to 35 PCR cycles using a proofreading high-fidelity polymerase and human adenomatous polyposis coli (APC) primer sets. From a 50 µl PCR reaction, 10 µl was analyzed on a 1% agarose gel. Lane 1, 1 kb ladder; lanes 2 and 3, 6.4 kb APC PCR product; lanes 4 and 5, 8.2 kb APC PCR product.

RNaseA Inhibition
 

 

 

RNase levels in different RNA samples. RNaseA, water, RT buffer (used to dilute RNaseA), spleen RNA (S RNA-1 and S RNA-2), and pancreas RNA (P RNA-1 and P RNA-2) from two different preparations were assayed for RNaseA activity using a commercially available kit. RNaseA (several fold higher than normally found in contaminated RNA preparations) was used as control to demonstrate varying levels of RNaseA activity in sample preparations.

 

RNase inhibition by the iScript™ cDNA synthesis kit. RNaseA, water, RT buffer (used to dilute RNaseA), and RNase + iScript cDNA kit were assayed for RNaseA activity using a commercially available kit. RNaseA (several fold higher than normally found in contaminated RNA preparations) was used as control to assess the effect of inhibition by the iScript cDNA kit.

 

Functional RT-qPCR assay demonstrating RNaseA inhibition by the iScript cDNA synthesis kit. 18S rRNA was amplified using iQ™ SYBR® Green supermix. Reverse transcription (RT) performed with iScript kit (); RT performed with spiked RNaseA (); RT performed with RNaseA but no RNase inhibitor (■). Two RNA inputs (1,000 and 0.1 pg) were reverse transcribed for all three groups. The cDNA synthesis reaction with RNase but no RNase inhibitor showed significant threshold cycle (CT) delay (3–5), whereas the iScript kit demonstrated powerful inhibition of RNaseA. 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

The iScript Select cDNA synthesis kit is a sensitive reverse transcription system offering flexible priming strategies. Unlike our iScript cDNA synthesis kit and iScript reverse transcription supermix for RT-qPCR, this kit comes with multiple reaction components (5x buffer, oligo[dT], random primers, proprietary gene-specific primer enhancer solution, and water) that must be combined during reaction setup.

Key Features and Benefits

  • Have flexibility in mRNA priming strategy — oligo(dT), random primer, and gene-specific primer enhancer solution provided in separate tubes
  • Synthesize long cDNAs up to 8 kb — quality controlled for reliable synthesis of cDNA >6 kb in length
  • Detect low-level target genes and conserve RNA during gene expression analysis — broad linear dynamic range of total input RNA (1 µg–1 pg) with a highly efficient RNase H+ MMLV reverse transcriptase
  • Avoid skewed data from RNA degradation — potent blend of RNaseA inhibitor protects RNA during setup and reverse transcription
Storage at –20°C Guaranteed for 12 months at –20°C in a constant temperature freezer (multiple freeze thaws not recommended)
iScript™ Select cDNA Synthesis Kit

170-8896
25 x 20 µl reactions, includes 5x reaction mix, iScript reverse transcriptase, oligo(dT), random primer mix, gene-specific primer enhancer solution, nuclease-free water

List Price:   $135.00
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iScript™ Select cDNA Synthesis Kit

170-8897
100 x 20 µl reactions, includes 5x reaction mix, iScript reverse transcriptase, oligo(dT), random primer mix, gene-specific primer enhancer solution, nuclease-free water

List Price:   $430.00
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iScript™ Select cDNA Synthesis Kit

170-8897BUN
500 x 20 µl reactions, includes 5x reaction mix, iScript reverse transcriptase, oligo(dT), random primer mix, gene-specific primer enhancer solution, nuclease-free water

List Price:   $2,035.00
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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) ]
10001023 Instruction Manual, iScript Select cDNA Synthesis Kit, Rev B Click to download
2894 iScript cDNA Synthesis Kits Flier, Rev F Click to download
5438 Convenient Multiplex PCR Assays With iQ Multiplex Powermix, Rev A Click to download