Reference genome-independent assessment of mutation density using restriction enzyme-phased sequencing.

ABSTRACT:

Background

The availability of low cost sequencing has spurred its application to discovery and typing of variation, including variation induced by mutagenesis. Mutation discovery is challenging as it requires a substantial amount of sequencing and analysis to detect very rare changes and distinguish them from noise. Also challenging are the cases when the organism of interest has not been sequenced or is highly divergent from the reference.

Results

We describe the development of a simple method for reduced representation sequencing. Input DNA was digested with a single restriction enzyme and ligated to Y adapters modified to contain a sequence barcode and to provide a compatible overhang for ligation. We demonstrated the efficiency of this method at SNP discovery using rice

SUBMITTER: Monson-Miller J 

PROVIDER: S-EPMC3305632 | biostudies-literature | 2012 Feb

REPOSITORIES: biostudies-literature