Bio-Rad has developed a system that combines reverse transcription with droplet digital PCR
The one-step RT-ddPCR kit for probes is designed to measure target RNA molecules with precision for applications such as gene expression analysis, miRNA analysis, and viral load quantitation.
Researchers can use the one-step RT-ddPCR kit 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 differences in gene expression levels, and determine copies of an RNA molecule without a standard curve.
The system 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.
