Random-primed, Phi29 DNA polymerase-based whole genome amplification

Curr Protoc Mol Biol. 2014 Jan 6:105:Unit 15.13.. doi: 10.1002/0471142727.mb1513s105.

Abstract

Whole-genome amplification by multiple displacement amplification (MDA) is a patented method to generate potentially unlimited genomic material when researchers are challenged with trace samples, or the amount of genomic DNA required for analysis exceeds the amount on hand. It is an isothermal reaction, using Phi29 DNA polymerase and random hexamer primers for unbiased amplification of linear DNA molecules, such as genomic DNA. The random-primed MDA reaction provides extensive amplification coverage of the genome, generates extremely long DNA products, and provides high DNA yields. This unit explains the reaction, and describes use of the commercial kits available.

Keywords: Phi29 DNA polymerase; isothermal amplification; multiple displacement amplification (MDA); whole-genome amplification (WGA).

MeSH terms

  • DNA / genetics*
  • DNA Primers / genetics*
  • DNA-Directed DNA Polymerase / metabolism*
  • Genome, Human / genetics*
  • Humans
  • Polymerase Chain Reaction / methods*

Substances

  • DNA Primers
  • DNA
  • DNA-Directed DNA Polymerase