Spacer-Based Macroarrays for CRISPR Genotyping

Methods Mol Biol. 2015:1311:111-31. doi: 10.1007/978-1-4939-2687-9_7.

Abstract

Macroarray-based analysis is a powerful and economic format to study variations in "clustered regularly interspaced short palindromic repeat (CRISPR)" loci in bacteria. To date, it was used almost exclusively for Mycobacterium tuberculosis and was named spoligotyping (spacer oligonucleotides typing). Here, we describe the pipeline of this approach that includes search of loci and selection of spacers, preparation of the membrane with immobilized probes and spoligotyping itself (PCR and reverse hybridization).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Corynebacterium diphtheriae / genetics
  • Genetic Loci / genetics
  • Genotyping Techniques / methods*
  • Membranes, Artificial
  • Oligonucleotide Array Sequence Analysis / methods*
  • Oligonucleotide Probes / genetics
  • Polymerase Chain Reaction

Substances

  • Membranes, Artificial
  • Oligonucleotide Probes