MetaHit Single Cell WGA

Abstract: In order to facilitate sequencing of uncultured phylotypes from the human gut microbiota it was first necessary to isolate individual bacterial cells via Laser microdissection microscopy. Isolated individual cells were lysed by heat treatment in a stabilising sample buffer. Genomic DNA released underwent Multiple Displacement Amplification (MDA) to amplify the whole genome, the Genomiphi V2 mini kit was used for this reaction. 16S rRNA gene PCR was used to determine the phylotype of the amplification. Samples of interest were submitted for illumina whole genome sequencing with the aim of generating the first genome sequence for the uncultured phylotypes isolated. The resulting sequence data will feed into the MetaHit consortium study.