Preparation of a 3-kb Mate-Pair Library for Illumina Sequencing

The "mate-pair" approach for dual end sequencing uses long library fragments (1000-10,000 bp) that are circularized by ligation to the ends of a single, common DNA adaptor of known sequence. Once the ends are mated to the adaptor, a final library of fragments that retain the adaptor and only the ends of the original, circularized piece of DNA can be obtained. This protocol describes how to prepare a 3-kb mate-paired-end library. The resulting amplified DNA is suitable for sequencing on the Illumina sequencer.