Improved genome recovery and integrated cell-size analyses of individual uncultured microbial cells and viral particles.

ABSTRACT: Microbial single-cell genomics can be used to provide insights into the metabolic potential, interactions, and evolution of uncultured microorganisms. Here we present WGA-X, a method based on multiple displacement amplification of DNA that utilizes a thermostable mutant of the phi29 polymerase. WGA-X enhances genome recovery from individual microbial cells and viral particles while maintaining ease of use and scalability. The greatest improvements are observed when amplifying high G+C content templates, such as those belonging to the predominant bacteria in agricultural soils. By integrating WGA-X with calibrated index-cell sorting and high-throughput genomic sequencing, we are able to analyze genomic sequences and cell sizes of hundreds of individual, uncultured bacteria, archaea, protist

SUBMITTER: Stepanauskas R 

PROVIDER: S-EPMC5519541 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature