Biotinylated amplicon sequencing: A method for preserving DNA samples of limited quantity.

ABSTRACT:

Background

Genomic testing is often limited by the exhaustible nature of human tissue and blood samples. Here we describe biotinylated amplicon sequencing (BAmSeq), a method that allows for the creation of PCR amplicon based next-generation sequencing (NGS) libraries while retaining the original source DNA.

Design and methods

Biotinylated primers for different loci were designed to create NGS libraries using human genomic DNA from cell lines, plasma, and formalin-fixed paraffin embedded (FFPE) tissues using the BAmSeq protocol. DNA from the original template used for each BAmSeq library was recovered after separation with streptavidin magnetic beads. The recovered DNA was then used for end-point, quantitative and droplet digital PCR (ddPCR) as well as NGS using a cancer gene

SUBMITTER: Cravero K 

PROVIDER: S-EPMC6104457 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature