Methods for Target Enrichment Sequencing via Probe Capture in Legumes

Methods Mol Biol. 2020:2107:199-231. doi: 10.1007/978-1-0716-0235-5_10.

Abstract

Target enrichment sequencing (TES) is a powerful approach to deep-sequencing the exome or genomic regions of interest with great depth. Although successfully and widely adopted in many plant species, TES is currently applied for genotyping of only a couple legume species. Here we describe an in-solution probe capture based method for application of TES in legumes. The topics cover probe design, library preparation, probe hybridization, as well as bioinformatic analysis for evaluation of target capture efficiency and identifying single nucleotide polymorphisms using generated sequencing data.

Keywords: Legume; Next generation sequencing; Probe design; Probe hybridization; Single nucleotide polymorphism; Target enrichment sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Computational Biology
  • Exome
  • Fabaceae / genetics*
  • Gene Expression Profiling / methods
  • Gene Library
  • Genotyping Techniques
  • High-Throughput Nucleotide Sequencing / methods*
  • Nucleic Acid Probes / genetics*
  • Polymorphism, Single Nucleotide*
  • Sequence Analysis, DNA / methods

Substances

  • Nucleic Acid Probes