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Detection and quantitative analysis of a specific RNA target e.g. for gene expression analyses can be performed via Reverse Transcription (RT) followed by real-time PCR (qPCR) or end-point PCR analysis (RT-qPCR or RT-PCR, respectively). RNA is first converted into complementary DNA (cDNA) by reverse transcriptase from total RNA (10 pg to 1 µg) or poly(A)-enriched RNA (e.g. mRNA, 1 pg to 20 ng). The cDNA is subsequently used as template for (q)PCR amplification of target specific fragments.

Optimized reaction mixes are available

  • for one-step cDNA synthesis and (q)PCR amplification (RT-(q)PCR) which minimizes the risk of contaminations and offers convenient handling especially for high throughput analyses.
  • with two read out options for qPCR amplification (ProbesMaster: based on dual-labeled (hydrolysis) probes, GreenMaster: based on dsDNA intercalator dye EvaGreen®).

EvaGreen® is a registered trademark and licensed for sale by Biotium, Inc., Hayward, CA, USA.