Rapid Gene Isolation Using MutChromSeq

Methods Mol Biol. 2017:1659:231-243. doi: 10.1007/978-1-4939-7249-4_20.

Abstract

MutChromSeq is an approach for isolation of genes and DNA sequences controlling gene expression in plants with complex and polyploid genomes. It involves a lossless complexity reduction by flow cytometric chromosome sorting and shotgun sequencing DNA from isolated chromosomes. Comparison of sequences from wild-type parental chromosome with chromosomes from multiple independently derived mutants identifies causative mutations in a single candidate gene or a noncoding sequence. MutChromSeq does not rely on recombination-based genetic mapping and does not exclude any DNA sequence from being targeted.

Keywords: Chromosome isolation; FISHIS; Flow cytometry and chromosome sorting; Gene cloning; Mutational genomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromosomes, Plant / genetics*
  • DNA, Plant / genetics*
  • Flow Cytometry / methods
  • Gene Expression Regulation, Plant
  • Genome, Plant
  • Genomics / methods*
  • In Situ Hybridization, Fluorescence / methods
  • Mutation*
  • Plants / genetics*
  • Polyploidy
  • Sequence Analysis, DNA / methods

Substances

  • DNA, Plant