A novel process of viral vector barcoding and library preparation enables high-diversity library generation and recombination-free paired-end sequencing.

ABSTRACT: Detailed characterization and mapping of oligonucleotide function in vivo is generally a very time consuming effort that only allows for hypothesis driven subsampling of the full sequence to be analysed. Recent advances in deep sequencing together with highly efficient parallel oligonucleotide synthesis and cloning techniques have, however, opened up for entirely new ways to map genetic function in vivo. Here we present a novel, optimized protocol for the generation of universally applicable, barcode labelled, plasmid libraries. The libraries are designed to enable the production of viral vector preparations assessing coding or non-coding RNA function in vivo. When generating high diversity libraries, it is a challenge to achieve efficient cloning, unambiguous barcoding and detailed charac

SUBMITTER: Davidsson M 

PROVIDER: S-EPMC5118689 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature