Whole Exome Library Construction for Next Generation Sequencing

  • Winnie S. LiangEmail author
  • Kristi Stephenson
  • Jonathan Adkins
  • Austin Christofferson
  • Adrienne Helland
  • Lori Cuyugan
  • Jonathan J. Keats
Part of the Methods in Molecular Biology book series (MIMB, volume 1706)


Whole exome sequencing (WES) is a DNA sequencing strategy that provides a survey of base substitutions across coding genomic locations and other regions of interest. As the coding portion of the genome encompasses only 1–2% of the entire genome, this approach represents a more cost-effective strategy to detect DNA alterations that may alter protein function, compared to whole genome sequencing. Although the research community has and is currently delineating the functional implications of sequence changes in noncoding regions of the genome, WES is a currently available assay that provides valuable information for both discovery research and precision medicine applications. In this chapter, we present a WES library preparation protocol using the KAPA Hyper Prep Kit with Agilent SureSelect Human All Exon V5+UTR probes that demonstrates high DNA-to-library conversion efficiency for sequencing on the Illumina HiSeq platform.

Key words

Whole exome sequencing Next generation sequencing DNA substitutions Rare diseases Coding region Library preparation 


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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Winnie S. Liang
    • 1
    Email author
  • Kristi Stephenson
    • 1
  • Jonathan Adkins
    • 1
  • Austin Christofferson
    • 1
  • Adrienne Helland
    • 1
  • Lori Cuyugan
    • 1
  • Jonathan J. Keats
    • 1
  1. 1.Translational Genomics Research Institute (TGen)PhoenixUSA

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