Extraction of Cell-Free DNA and Subsequent Preparation of NGS Libraries in a High-Throughput, Cost-Effective Manner

摘要

An efficient workflow from sample to sequencer is critical to maintaining integrity of circulating, cell-free DNA (cfDNA) from liquid biopsy samples. Each step in the process; blood collection, DNA extraction, and library creation, can affect the quality of sequencing results. Here we discuss cfDNA isolation on the PerkinElmer chemagicTM 360 system, followed by use of the PerkinElmer Sciclone? NGSx to generate next generation sequencing (NGS) libraries from cfDNA using the Accel-NGS? 2S PCR-Free Library kit from Swift Biosciences. cfDNA was extracted from plasma collected from whole blood samples utilizing the chemagic 360 instrument and chemistry, enabling automated isolation of high quality samples. Multiple methods were applied to check for contamination of high molecular weight DNA and integrity of the cfDNA.