Conversion of a Fosmid library to an Illumina-compatible Fosill jumping library. (A,B) The two Nb.BbvCI sites in the vector are nicked. (C) The nicks are translated in opposite directions into the cloned insert. (D) The insert is cleaved at the two translated nicks as well as at nicks originating at any BbvCI sites within the genomic DNA sequence. (E) Fragments are circularized by intramolecular ligation. (F) Recircularized vector molecules serve as templates for inverse PCR with full-length Illumina enrichment primers that include the sequences required for bridge-amplification and paired-end sequencing of the coligated termini of the original Fosmid insert on the Illumina flow cell.
