GSM5049798: Sensitivity_Testing_MIPP-seq2; Homo sapiens; OTHER
MIPP-Seq: Unique primers are designed to prevent sharing binding sites and avoiding known mutations in the individual and general population but are within 250 nucleotides of the target mutation. Once designed, unique 10nt barcodes are appended to the reverse primer, along with the standard Ion Torrent platform specific adapters.  The primers designed above amplify targets using reduced cycling  (20cycles) with a high-fildelity enzyme and minimal amount of DNA (25-50ng). As all primers are unique and can be identified using the primer sequence, recations from up to 96 differnet mutations are pooled and PCR purified, before being diluted to 100pm for sequencing on the Ion Torrent S5 with 400nt reads platform for ultra-deep coverage.