This video demonstrates the multiplex droplet polymerase chain reaction technique for the simultaneous quantification of methylated DNA regions in different white blood cell populations as an early biomarker for clinical diagnosis and prognosis.
Protocol
1. Droplet generation and PCR NOTE: Table 1 outlines information on the forward and reverse primers along with the double-quenched hydrolysis probes for C-LESS, CD3Z, and Foxp3 genes, which are required for the multiplex amplification of demethylated gene targets. Prepare the master mix as described in Table 2. Thaw all components of the master mix except for the enzyme mix. Mix the master mix thoroughly by pipetting up-d…