Knowledge-Based Identification of Plant Stress Regulators

Here we adapt digital droplet PCR to operate under COLD-PCR conditions, thereby preferentially amplifying mutation-containing sequences in droplets.  COLD-PCR enables amplicons with mutations at any position to be amplified preferentially, while wild-type samples are preferentially suppressed. The preferentially amplified sequences can subsequently be sequenced. We validate the merging of technologies in serially diluted TP53 and KRAS mutated sequences. By merging COLD-PCR and digital -PCR technologies we enable digital mutation scanning, a new frontier for digital-PCR. A single reaction can now interrogate numerous mutations, and replaces multiple individual digital-PCR reactions.