Mate pair sequencing involves generating long-insert paired-end DNA libraries useful for a number of sequencing applications, including:
Combining data generated from mate pair library sequencing with that from short-insert paired-end reads provides a powerful combination of read lengths for maximal sequencing coverage across the genome.
Following DNA fragmentation, the DNA fragments are end-repaired with labeled dNTPs. The DNA fragments are circularized, and non-circularized DNA is removed by digestion. Circular DNA is fragmented, and the labeled 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 for paired-end cluster generation, followed by sequencing utilizing an Illumina next-generation sequencing (NGS) system.
Efficient protocol enables the highest genomic diversity of any next-generation platform.
Simple workflow with limited hands-on time and multiple stopping points.
Requires as little as 1 μg of starting material.