Rapid multi-locus sequence typing using microfluidic biochips.

ABSTRACT:

Background

Multiple locus sequence typing (MLST) has become a central genotyping strategy for analysis of bacterial populations. The scheme involves de novo sequencing of 6-8 housekeeping loci to assign unique sequence types. In this work we adapted MLST to a rapid microfluidics platform in order to enhance speed and reduce laboratory labor time.

Methodology/principal findings

Using two integrated microfluidic devices, DNA was purified from 100 Bacillus cereus soil isolates, used as a template for multiplex amplification of 7 loci and sequenced on forward and reverse strands. The time on instrument from loading genomic DNA to generation of electropherograms was only 1.5 hours. We obtained full-length sequence of all seven MLST alleles from 84 representing 46 different Sequen

SUBMITTER: Read TD 

PROVIDER: S-EPMC2868872 | biostudies-literature | 2010 May

REPOSITORIES: biostudies-literature