Protocol

Whole-Genome Sequencing: Automated, Indexed Library Preparation

  1. W. Richard McCombie

Abstract

This protocol describes an automated procedure for constructing an indexed Illumina DNA library. With this method, genomic DNA fragments are produced by sonication, using high-frequency acoustic energy to shear DNA. Double-stranded DNA (dsDNA) will fragment when exposed to the energy of adaptive focused acoustic shearing (AFA). The resulting DNA fragments are ligated to adaptors, amplified by polymer chain reaction (PCR), and subjected to size selection using magnetic beads. The product is suitable for use as template in whole-genome sequencing.

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.prot094649 Cold Spring Harb Protoc 2017: pdb.prot094649- © 2017 Cold Spring Harbor Laboratory Press
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