De novo assembly and next-generation sequencing to analyse full-length gene variants from codon-barcoded libraries.

ABSTRACT: Interpreting epistatic interactions is crucial for understanding evolutionary dynamics of complex genetic systems and unveiling structure and function of genetic pathways. Although high resolution mapping of en masse variant libraries renders molecular biologists to address genotype-phenotype relationships, long-read sequencing technology remains indispensable to assess functional relationship between mutations that lie far apart. Here, we introduce JigsawSeq for multiplexed sequence identification of pooled gene variant libraries by combining a codon-based molecular barcoding strategy and de novo assembly of short-read data. We first validate JigsawSeq on small sub-pools and observed high precision and recall at various experimental settings. With extensive simulations, we then apply Jigs

SUBMITTER: Cho N 

PROVIDER: S-EPMC4595759 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature