iScript™ cDNA Synthesis Kit

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Overview

Broad Dynamic Range

The iScript™ cDNA synthesis kit performs across a broad range of concentrations. Input RNA (amounts shown in inset) was reverse transcribed and the resulting cDNA was amplified using iQ™ SYBR® Green supermix. The iScript cDNA synthesis kit performs over six orders of magnitude with high sensitivity. Standard curve r = 0.998, efficiency = 96.5%, RFU, relative fluorescence units.

Excellent Sensitivity

Sensitivity of the iScript™ cDNA synthesis kit in RT-PCR using low amounts of input RNA. cDNA was synthesized at 42°C and amplified with iTaq™ DNA polymerase using β-actin primers. M, EZ Load™ 100 bp molecular ruler; C, no-template negative control.

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 iScript™ One-Step RT-PCR Kit With SYBR® Green iScript™ One-Step RT-PCR Kit for Probes

The iScript cDNA synthesis kit is a sensitive and easy-to-use first-strand cDNA synthesis kit for gene expression analysis using real-time qPCR. This two-tube kit is optimized to yield sensitive, unbiased representation over a broad dynamic range, with minimal setup and reaction time.

Key Features and Benefits

  • Have ease of use and consistency — 2-tube (iScript reaction mix and iScript reverse transcriptase) format for faster setup
  • 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
  • Utilize dilute RNA samples — reaction mix and iScript enzyme blend format allows up to 15 µl of RNA sample in a 20 µl cDNA synthesis reaction
  • Prevent 5' and 3' bias of your target genes and design primers in any region — optimum blend of oligo(dT) and random primers in the 5x supermix
  • Avoid skewed data from RNA degradation — potent blend of RNaseA inhibitor protects RNA during setup and reverse transcription
  • Complete cDNA synthesis and qPCR on the same day — fast 40 min cDNA synthesis protocol
Storage at –20°C Guaranteed for 12 months at –20°C in a constant temperature freezer (multiple freeze thaws not recommended)
iScript™ cDNA Synthesis Kit

170-8890
25 x 20 µl reactions, includes 5x reverse-transcription reaction mix, iScript reverse transcriptase, nuclease-free water

List Price:   $149.00
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iScript™ cDNA Synthesis Kit

170-8891
100 x 20 µl reactions, includes 5x reverse-transcription reaction mix, iScript reverse transcriptase, nuclease-free water

List Price:   $446.00
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iScript™ cDNA Synthesis Kit

170-8891BUN
500 x 20 µl reactions, includes 5x reverse-transcription reaction mix, iScript reverse transcriptase, nuclease-free water

List Price:   $2,065.00
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