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HIGH SENSITIVITY qRT-PCR

MessageSensor Reverse Transcription Kit For One Step qRT-PCR


More sensitive than SuperScript II and other RNase H MMLV reverse transcriptases

Linear quantitation of mRNA targets over a 106-fold range

Quantitate transcripts from as little as 500 fg total RNA 1/20th of a single cell!

Fast, flexible RT step complete in 15 min

Use reaction temperatures up to 50C

Compatible with TaqMan, SYBR Green, and gel-based detection methods


RNase H RT Reduces Detection Sensitivity in RT-PCR
RNase H minus (RNase H) reverse transcriptases (RTs) are a popular choice among researchers for applications that require a cDNA synthesis step. RNase H RTs are available from several manufacturers, with SuperScript II (Invitrogen) being the most widely used. RNase H RTs can generate longer cDNA products with higher yields than wild type (RNase H+) RTs, which are important criteria when generating cDNA libraries or performing DNA microarray analysis. However, these benefits are unnecessary for qRT-PCR where amplicons average 100 bp. Additionally, RNase H RT can limit the sensitivity of qRT-PCR detection (Figures 1 and 2). If the RNA template is not degraded after first strand cDNA synthesis, it can bind to the newly-synthesized cDNA and restrict its accessibility to primers during subsequent PCR amplification.
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Related biology technology :

1. Designing a Successful qRT-PCR Experiment
2. Isolate High Quality Total RNA from LCM Samples Suitable for Microarray and qRT-PCR Analysis
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