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