Illumina sequencing of 15 deafness genes using fragmented amplicons.

ABSTRACT:

Background

Resequencing of deafness related genes using GS FLX massive parallel sequencing of PCR amplicons spanning selected genes has previously been reported as a successful strategy to discover causal variants. The amplicon lengths were designed to be smaller than the sequencing read length of GS FLX technology, but are longer than Illumina sequencing technology read lengths. Fragmentation is thus required to sequence these amplicons using high throughput Illumina technology.

Methods

We performed Illumina sequencing in 4 patients on 563 multiplexed amplicons covering the exons of 15 genes involved in the hearing process. After exploring several fragmentation strategies, the amplicons were fragmented using Covaris sonication prior to library preparation. CLC genomic workb

SUBMITTER: Van Nieuwerburgh F 

PROVIDER: S-EPMC4266979 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature