Amplification testing in breast cancer by multiplex ligation-dependent probe amplification of microdissected tissue

Cathy B. Moelans, Roel A. De Weger, Paul J. Van Diest

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

This chapter describes a method for the rapid assessment of gene copy numbers in laser-microdissected materials using multiplex ligation-dependent probe amplification (MLPA). An MLPA is a powerful multiplex PCR technique that can identify gains, amplification, or losses of up to 50 genes in a single experiment, thereby requiring only minute quantities of DNA extracted from frozen or paraffin-embedded materials. A previous study in breast cancer has shown that MLPA can detect amplifications in cases with a tumor percentage lower than 10%, but still a low tumor percentage in the tissue tested could obscure low levels of amplification due to dilution of the tumor cell population by normal cells. Laser capture microdissection allows enrichment of tumor cells by eliminating background noise from normal and preinvasive cells, thereby increasing specificity and sensitivity. This chapter describes a method for MLPA analysis using invasive breast tumor cells acquired by laser capture microdissection. This protocol can also be applied to MLPA analysis of preinvasive lesions and metastases.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages107-118
Number of pages12
Volume755
DOIs
Publication statusPublished - 2011
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume755
ISSN (Print)10643745

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Keywords

  • Cancer
  • Coffalyser
  • Laser microdissection
  • MLPA
  • Multiplex ligation-dependent probe amplification

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Moelans, C. B., De Weger, R. A., & Van Diest, P. J. (2011). Amplification testing in breast cancer by multiplex ligation-dependent probe amplification of microdissected tissue. In Methods in Molecular Biology (Vol. 755, pp. 107-118). (Methods in Molecular Biology; Vol. 755). https://doi.org/10.1007/978-1-61779-163-5_9