Targeted Locus Amplification and Next-Generation Sequencing

Quint P Hottentot; M van Min; E Splinter; Stefan J White

doi:10.1007/978-1-4939-6442-0_13

Targeted Locus Amplification and Next-Generation Sequencing

Methods Mol Biol. 2017:1492:185-196. doi: 10.1007/978-1-4939-6442-0_13.

Authors

Quint P Hottentot¹, M van Min², E Splinter³, Stefan J White⁴

Affiliations

¹ Leiden Genome Technology Center, Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
² Cergentis, Utrecht, The Netherlands.
³ Cergentis, Utrecht, The Netherlands. erik.splinter@cergentis.com.
⁴ Leiden Genome Technology Center, Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. s.j.white@lumc.nl.

PMID: 27822865
DOI: 10.1007/978-1-4939-6442-0_13

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.

Keywords: Copy number variation; Next-generation sequencing; Structural variation; Variant detection.

MeSH terms

DNA Primers
High-Throughput Nucleotide Sequencing / methods*
Polymerase Chain Reaction / methods*

Substances

DNA Primers