Targeted Locus Amplification and Haplotyping

Juliet W. Lefferts, Vera Boersma, Marne C. Hagemeijer, Karima Hajo, Jeffrey M. Beekman, Erik Splinter*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

3 Citations (Scopus)
1 Downloads (Pure)

Abstract

Targeted locus amplification (TLA) allows for the detection of all genetic variation (including structural variation) in a genomic region of interest. As TLA is based on proximity ligation, variants can be linked to each other, thereby enabling allelic phasing and the generation of haplotypes. This allows for the study of genetic variants in an allele-specific manner. Here, we provide a step-by-step protocol for TLA sample preparation and a complete bioinformatics pipeline for the allelic phasing of TLA data. Additionally, to illustrate the protocol, we show the ability of TLA to re-sequence and haplotype the complete cystic fibrosis transmembrane (CFTR) gene (> 200 kb in size) from patient-derived intestinal organoids.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages31-48
Number of pages18
Volume2590
DOIs
Publication statusPublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2590
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cystic fibrosis
  • Genetic variation
  • Haplotyping
  • Next generation sequencing
  • Organoids
  • Phasing
  • Proximity ligation
  • Targeted Locus Amplification (TLA)

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