Pacific Biosciences of California Inc (PACB)

[Paulieme]

Fosmid-Based ResequencingThe resequencing of some genomic regions–including the MHC and KIR regions–has been time consuming and expensive due to their size and repetitive structure. Our method combines a fosmid-based approach with next generation sequencing to provide you with complete, high fidelity, and phased sequence.

Fosmid libraries are constructed from fragmented genomic DNA and are diluted into 96-well plates to reduce the complexity of each well. The use of fosmids provides the advantage of a higher scale order on the highly repetitive structures of complex genomic regions.

Next generation sequencing, using the Illumina MiSeq Sequencing System allows us to identify target fosmids within each well in a single step. Target fosmids are isolated from wells via recombineering and sequenced together in a single sequencing run using either the MiSeq or PacBio’s Single Molecule Real-Time (SMRT®) long-read sequencing technology. Fosmid sequences are assembled using overlaps, producing phased, high quality data on the megabase scale.

(click on link for charts) http://www.sciscogenetics.com/technology/fosmid-based-resequencing/