Multiplexed Mutation Detection in Circulating Cell Free DNA and FFPE Samples

摘要

Detection of somatic mutations in oncogenes is challenging since percent tumor content in clinical samples is variable and is compounded by tumor heterogeneity. Additionally, circulating cfDNA and FFPE samples are typically limited in quantity, and FFPE samples can also be highly damaged. To address these challenges, we developed a single tube, multiplexed amplicon sequencing method that employs hundreds of primer pairs for amplification of target loci, producing ready-to-run libraries for Illumina sequencing. The simple two-step workflow takes two hours: multiplexed PCR followed by a 10 minute adapter ligation.