An efficient pipeline for the generation and functional analysis of human BRCA2 variants of uncertain significance.

ABSTRACT: The implementation of next-generation sequence analysis of disease-related genes has resulted in an increasing number of genetic variants with an unknown clinical significance. The functional analysis of these so-called "variants of uncertain significance" (VUS) is hampered by the tedious and time-consuming procedures required to generate and test specific sequence variants in genomic DNA. Here, we describe an efficient pipeline for the generation of gene variants in a full-length human gene, BRCA2, using a bacterial artificial chromosome. This method permits the rapid generation of intronic and exonic variants in a complete gene through the use of an exon-replacement strategy based on simple site-directed mutagenesis and an effective positive-negative selection system in E. coli. The func

SUBMITTER: Hendriks G 

PROVIDER: S-EPMC6706860 | biostudies-literature | 2014 Nov

REPOSITORIES: biostudies-literature