Genomic DNA library preparation for resistance gene enrichment and sequencing (RenSeq) in plants

Methods Mol Biol. 2014:1127:291-303. doi: 10.1007/978-1-62703-986-4_22.

Abstract

Enrichment of genomic DNA for genes of interest prior to high-throughput sequencing offers an efficient and cost-effective approach to reduce genome complexity. Target enrichment typically yields higher read-depth for selected genes and is therefore suitable for determination of sequence polymorphisms and enables multiplexing of samples. Target enrichment also provides a means to annotate specific gene families within the sequenced organisms without the requirements for gene models. Here we describe enrichment procedures for NB-LRR-type plant resistance genes that can, for example, be used to establish the NB-LRR gene complements of individual plants and to map resistances more rapidly using a bulked segregant analysis.

MeSH terms

  • DNA Fragmentation
  • DNA, Plant / isolation & purification
  • DNA, Plant / metabolism*
  • Disease Resistance / genetics*
  • Genes, Plant*
  • Genomic Library*
  • Plant Diseases / genetics*
  • Plant Diseases / microbiology*
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA / methods*

Substances

  • DNA, Plant