High-throughput fosmid sequencing

Hits isolated from our screening program must be sequenced to help identify the gene(s) responsible for the activity of interest. To that end, we routinely sequence fosmid DNA. We are developing simple and effective methods for high-throughput fosmid sequencing that use isothermal amplification of fosmid DNA directly from lysed cells for sequencing library sample preparation. Currently, up to 96 multiplexed fosmids can be analyzed in a single sequencing run. Our fosmid sequencing efforts strongly support our screening program, helping us to speed-up gene discovery.

Example of a fosmid sequence map.
The map of a 41 kb fosmid DNA sequence is shown. ORFs were predicted using MetaGeneMark and represented here with arrows.