Whole-Genome Sequencing and Assembly with High-Throughput, Short-Read Technologies (original) (raw)
Figure 4
Implementation of sequencing protocol.
The genome is first fragmented into 150 Kb pieces, of which we randomly select 200,000. Each piece is individually cloned and further fragmented into small pieces suitable for sequencing. We then ligate sequencing adapters that include a 5-base tag that is unique to each clone within a 266-clone “batch”. After amplifying the fragments on beads, a batch of 266 clones are multiplexed together on a 400,000 read plate, and the first five bases of each read are used to identify the source clone. By running 750 plates in this fashion, we can fully sequence a mammalian genome to 20.0x coverage.