Jiali Gu, Sheila Rochette, PhD, Joy Nakitandwe, PhD, St. Jude Children’s Research Hospital

Validating the Genetic Mutations Identified by the PCGP

The PCGP Validation Laboratory was established to independently confirm the genetic lesions identified by whole-genome sequencing and analyze the mutation frequencies in expanded validation cohorts. Four sequencing methods—Sanger fluorescent capillary-electrophoresis sequencing, pyrosequencing, sequencing by synthesis, and single-molecule sequencing—have been evaluated during the course of the project.

To date, more than 500,000 putative lesions have been sequenced and verified by the PCGP Validation Laboratory. Improvements in protocols and technologies have increased the lab’s sequencing capacity from 60 gigabases of DNA sequence per week in 2010 to 1.2 terabases per week in 2012. This increased sequencing capacity has also allowed the laboratory to perform whole-exome sequencing and RNA-sequencing on almost 2000 patients in the validation cohorts.