A Simple Method for Obtaining Absolute Quantification of DNA Molecules Using the Innovative Droplet Digital PCR Technology
Droplet digital PCR is considered the third generation of PCR technology. This technology enables the absolute measure of target DNA molecules, rather than the relative measure obtained from real-time PCR. Absolute quantification is advantageous in several applications, such as measuring copy number variation, detecting rare sequences and mutations, and analyzing gene expression.
Bio-Rad's QX100™ Droplet Digital™ PCR (ddPCR™) system provides a revolutionary approach to target DNA quantification with a simple and elegant workflow. The QX100 droplet generator partitions samples into thousands of nanoliter-sized droplets. After PCR on a thermal cycler, such as C1000 Touch™ thermal cycler, droplets from every sample are streamed single file in the QX100 droplet reader. Each droplet provides a fluorescent positive or negative signal indicating whether the target DNA is present or not. Positive and negative droplets are counted and the concentration of target DNA is automatically calculated by the software.
Step by Step with the QX100 ddPCR System
The experimental workflow, depicted in the video below, begins with PCR-ready samples and consists of the following steps: generating droplets, performing PCR, reading droplets, and analyzing the results. Steps involve minimal hands-on time and the procedures are easy to perform.