Targeted Locus Amplification and Next-Generation Sequencing.
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.
Leiden Genome Technology Center, Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
Quint P
QP
Hottentot
Cergentis, Utrecht, The Netherlands.
M
M
van Min
Cergentis, Utrecht, The Netherlands. erik.splinter@cergentis.com.
E
E
Splinter
Leiden Genome Technology Center, Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. s.j.white@lumc.nl.
Stefan J
SJ
White
D017931:DNA Primers; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers
Copy number variation; Next-generation sequencing; Structural variation; Variant detection