Protocol

Whole-Genome Sequencing: Automated, Nonindexed Library Preparation

  1. W. Richard McCombie

Abstract

This protocol describes an automated procedure for constructing a nonindexed Illumina DNA library and relies on the use of a CyBi-SELMA automated pipetting machine, the Covaris E210 shearing instrument, and the epMotion 5075. With this method, genomic DNA fragments are produced by sonication, using high-frequency acoustic energy to shear DNA. Here, double-stranded DNA is fragmented when exposed to the energy of adaptive focused acoustic shearing (AFA). The resulting DNA fragments are ligated to adaptors, amplified by polymerase 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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This Article

  1. Published in Advance November 1, 2016, doi: 10.1101/pdb.prot094623 Cold Spring Harb Protoc 2017: pdb.prot094623- © 2017 Cold Spring Harbor Laboratory Press
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