A Resource for Life Science Research

A New Paradigm for Precise Quantitation of RNA

By BioRadiations Staff

The one-step RT-ddPCR kit for probes creates a new paradigm for the precise quantitation of RNA by combining reverse transcription with Droplet Digital™ PCR (ddPCR™), providing an absolute measure of target RNA molecules with unrivaled precision and sensitivity. The kit is formulated for efficient and sensitive reverse transcription over a wide linear dynamic range of input RNA for use in ddPCR. The one-step RT-ddPCR kit for probes enables researchers to determine actual copy numbers of target RNA across samples as opposed to relative measurements determined by real-time PCR.

In combination with the QX100™ Droplet Digital™ PCR system, the one-step RT-ddPCR kit for probes lets you:

  • Enrich for rare target RNA sequences
  • Detect small differences in gene expression levels
  • Determine the number of copies of an RNA molecule without a standard curve

Key Features and Benefits

  • Unique hot-start reverse transcriptase enables convenient reaction setup
  • Reverse transcription reaction is performed at 55–60°C, which enhances the specificity and efficiency of primer-mediated cDNA conversion
  • Thermostable enzymes allow RNA template to be reverse transcribed and subsequently amplified in the same reaction tube
  • Template-specific primer annealing at elevated temperatures significantly improves stringency and melting of secondary structures
ddPCR absolute quantification of EEF2 and GAPDH gene transripcts using the one-step RT-ddPCR kit for probes


Fig 1. ddPCR absolute quantification of EEF2 (A) and GAPDH (B) gene transcripts using the one-step RT-ddPCR kit for probes. Raji total RNA was serially diluted twofold from 5 ng to 310 fg per 20 μl reaction. The data demonstrate the precision and sensitivity of the QX100 Droplet Digital PCR system in detecting thousands of copies down to a single copy per microliter, N = 6. NTC = no template control.
 

Ordering Information

Catalog #Product Description

186-3021

One-Step RT-ddPCR Kit for Probes, 2 x 1 ml, 200 x 20 μl reactions, 2x RT-ddPCR mix, contains manganese acetate

186-3022

One-Step RT-ddPCR Kit for Probes, 5 x 1 ml, 500 x 20 μl reactions, 2x RT-ddPCR mix, contains manganese acetate

186-3001QX100 Droplet Digital PCR system with computer, includes droplet generator, droplet reader, laptop computer, software, and associated component consumables

Practice of the polymerase chain reaction (PCR) may require a license.