Whole genome amplification and de novo assembly of single bacterial cells.

ABSTRACT:

Background

Single-cell genome sequencing has the potential to allow the in-depth exploration of the vast genetic diversity found in uncultured microbes. We used the marine cyanobacterium Prochlorococcus as a model system for addressing important challenges facing high-throughput whole genome amplification (WGA) and complete genome sequencing of individual cells.

Methodology/principal findings

We describe a pipeline that enables single-cell WGA on hundreds of cells at a time while virtually eliminating non-target DNA from the reactions. We further developed a post-amplification normalization procedure that mitigates extreme variations in sequencing coverage associated with multiple displacement amplification (MDA), and demonstrated that the procedure increased sequencing effi

SUBMITTER: Rodrigue S 

PROVIDER: S-EPMC2731171 | biostudies-literature | 2009 Sep

REPOSITORIES: biostudies-literature