Protocol

PCR Protocols

Volume 687 of the series Methods in Molecular Biology pp 193-205

Date:

The Many Faces of MLPA

  • Thomas OhnesorgAffiliated withMolecular Development, Murdoch Children’s Research Institute, Royal Children’s Hospital
  • , Erin TurbittAffiliated withMolecular Development, Murdoch Children’s Research Institute, Royal Children’s Hospital
  • , Stefan J. WhiteAffiliated withMolecular Development, Murdoch Children’s Research Institute, Royal Children’s Hospital

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Abstract

Multiplex Ligation-dependent Probe Amplification (MLPA) is a PCR-based technique that was developed for identifying deletions and duplications in genomic DNA. The simplicity and sensitivity of this approach has led to it being implemented in many laboratories around the world. Since the original publication, there have been several variants of MLPA described, allowing the quantitative analysis of mRNA transcript levels, CpG methylation, complex genomic regions, and DNaseI hypersensitive sites. This chapter outlines the basic MLPA protocol, describes the different modifications and applications that have been published, and discusses the critical points during each of the steps.

Key words

MLPA Copy number variation Deletion Duplication Mutation screening Methyla­tion Expression analysis DNaseI hypersensitivity