iScript™ One-Step RT-PCR Kit for Probes

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Overview

Broad Dynamic Range
Broad Dynamic Range

The iScript™ one-step RT-PCR kit for probes delivers unparalleled results over an extremely wide dynamic range. RNA (1 µg to 100 fg) isolated from HeLa cells using the Aurum™ total RNA kit was reverse-transcribed and amplified using primers to β-actin and a FAM-labeled detection probe. Each dilution was performed in triplicate, and RT-PCR was carried out on the iCycler iQ® real-time detection system. Standard curve r = 1.000, slope = –3.39, and efficiency = 97.2%, RFU, relative fluorescence units.

Robust Duplex RT-qPCR
Robust Duplex RT-qPCR

The iScript™ one-step RT-PCR kit for probes yields excellent results in duplex real-time reaction. Two gene targets from HeLa total RNA were reverse-transcribed and amplified in duplex over a 10-fold serial dilution using probes and primers to tubulin and β-actin. Each dilution was performed in triplicate, and the RT-PCR was carried out on the iCycler iQ® real-time detection system. A dual-labeled FAM probe was used to detect the β-actin gene product (top panel) in the first dye layer. Standard curve r = 1.000, slope = -3.53, and efficiency = 92%. In the second dye layer, a dual-labeled Cy5 probe was used to detect the tubulin gene product (lower panel). Standard curve r = 1.000, slope = -3.27, and efficiency = 102%, RFU, relative fluorescence units.

iScript™ Kit Selector
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™ one-step RT-PCR kit for probes is a highly sensitive solution for one-step real-time quantitative RT-PCR on a broad range of thermal cyclers. cDNA synthesis and PCR amplification are conveniently carried out in the same tube, using the powerful combination of iScript RNase H+ MMLV reverse transcriptase and hot-start iTaq™ DNA polymerase.

Key Features and Benefits

  • Maximize throughput and reduce the contamination risk of RT-qPCR analysis — 1-step RT-qPCR protocol allows quick and easy screening of a large number of samples
  • 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-RT
  • Benefit from superior compatibility with hybridization probe chemistry — optimized for sensitive detection using all types of hybridization probes, including dual-labeled oligonucleotide probes, FRET probes, and molecular beacons
  • Prevent nonspecific amplification and primer dimers — iTaq antibody-mediated hot-start DNA polymerase contains an antibody that sequesters activity prior to the initial PCR denaturation step
  • 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™ One-Step RT-PCR Kit for Probes

170-8894
50 x 50 μl reactions, one-step real time RT-PCR kit, includes iScript reverse transcriptase, 2x probes RT-PCR reaction mix, nuclease-free water

List Price:   $185.00
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iScript™ One-Step RT-PCR Kit for Probes

170-8895
200 x 50 μl reactions, one-step real-time RT-PCR kit, includes iScript reverse transcriptase, 2x probes RT-PCR reaction mix, nuclease-free water

List Price:   $675.00
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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 ]
4106270 Instruction Manual, iScript One-Step RT-PCR Kit for Probes, Rev C Click to download
3066 iScript One-Step RT-PCR Kit Flier, Rev C Click to download [ Add to Cart ]
5438 Convenient Multiplex PCR Assays With iQ Multiplex Powermix, Rev A Click to download
5962 Fast Optimization of a Multiplex Influenza Identification Panel Using a Thermal Gradient, Rev A Click to download [ Add to Cart ]