The one-step RT-ddPCR kit for probes, introduced by Bio-Rad Laboratories Inc., provides researchers with the ability to measure target RNA molecules with precision and sensitivity for applications such as gene expression analysis, miRNA analysis, and viral load quantitation.
The one-step RT-ddPCR kit creates a new paradigm for the precise quantitation of RNA by combining reverse transcription with droplet digital PCR. Researchers can use the one-step RT-ddPCR kit for probes with Bio-Rad’s QX100 Droplet Digital PCR system to determine small differences in copy number of target RNA that they were previously unable to measure using real-time PCR.
In combination with the QX100 ddPCR system, the one-step RT-ddPCR kit for probes allows scientists to:
• Enrich for rare target RNA sequences
• Detect small differences in gene expression levels
• Determine copies of an RNA molecule without a standard curve
The one-step RT-ddPCR kit for probes follows the ddPCR workflow except it partitions sample RNA instead of DNA or cDNA. The sample is partitioned into 20,000 droplets, with target and background RNA randomly distributed among them. After reverse transcription, the resulting cDNA is PCR amplified for target detection. Each droplet delivers a positive or negative fluorescent signal, indicating whether target RNA is present or absent. Positive and negative droplets are counted and software calculates the concentration of target RNA in copies per microliter.