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Genotyping pp 185-196 | Cite as

Targeted Locus Amplification and Next-Generation Sequencing

  • Quint P. Hottentot
  • M. van Min
  • E. Splinter
  • Stefan J. White
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1492)

Abstract

Despite developments in targeted and whole-genome sequencing, the robust detection of all genetic variation, including structural variants, in and around genes of interest and in an allele-specific manner remains a challenge. Targeted locus amplification (TLA) is a cross-linking-based technique that generates complex DNA libraries covering >100 kb of contiguous sequence surrounding one primer pair complementary to a short locus-specific sequence. In combination with next-generation sequencing, TLA enables the complete sequencing and haplotyping of targeted regions of interest. Here we outline the basis of TLA, together with a detailed protocol of the technique.

Key words

Variant detection Structural variation Copy number variation Next-generation sequencing 

Notes

Acknowledgements

We thank Marieke Simonis for data analysis.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Leiden Genome Technology Center, Department of Human GeneticsLeiden University Medical CenterLeidenThe Netherlands
  2. 2.CergentisUtrechtThe Netherlands

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