The Mate Pair Library Preparation Kit provides reagents needed to generate 400-600 bp libraries with inserts from 2-5 kb in size. Prepared libraries consist of short fragments made up of two segments that were originally separated by several kilobases in the genome. Mate Pairs are sequenced using a two-adapter strategy identical to the paired-end sequencing method.
Following DNA fragmentation, 2-5 Kb fragments are end-repaired with biotin labeled dNTPs. The DNA fragments are circularized, and non-circularized DNA is removed by digestion. Circular DNA is fragmented and biotin labels of the fragments (corresponding to the ends of the original DNA ligated together) are affinity purified. Purified fragments are end-repaired and ligated to Illumina Paired-End sequencing adapters.
Additional sequences complementary to the flow cell oligonucleotides are added to the adapter sequence with tailed PCR primers. The final prepared libraries consist of short fragments made up of two DNA segments that were originally separated by several kilobases. These libraries are ready to load on cBot or the Cluster Station for cluster generation.