Protocol

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

  1. W. Richard McCombie

Abstract

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.

Footnotes

  • From the Molecular Cloning collection, edited by Michael R. Green and Joseph Sambrook.

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  1. Published in Advance November 1, 2016, doi: 10.1101/pdb.prot094656 Cold Spring Harb Protoc © 2017 Cold Spring Harbor Laboratory Press
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