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Detection of Copy Number Changes at Multiple Loci in DNA Prepared from Formalin-Fixed, Paraffin-Embedded Tissue by Multiplex Ligation-Dependent Probe Amplification

  • Minoru Takata
Part of the Methods in Molecular Biology™ book series (MIMB, volume 439)

Abstract

With the increasing knowledge on the genetic alterations associated with various cancers, molecular analysis of these alterations on formalin-fixed paraffin-embedded tissue is of increasing importance. Multiplex ligation-dependent probe amplification (MLPA) is a novel technique to measure the copy number of up to 45 nucleic acid sequences in a single reaction. This method relies on sequence-specific probe hybridization to genomic DNA followed by multiplex-PCR amplification of the hybridized probe, and semi-quantitative analysis of the resulting PCR products. This method is easy to perform; requiring as little as 50 ng of DNA extracted from formalin-fixed, paraffin-embedded (FFPE) tissue.

Keywords

Chromosome aberrations FFPE tissue DNA imbalances melanocytic neoplasm 

References

  1. 1.
    1. Schouten JP, McElgunn CJ, Waaijer R, Zwijnenburg D, Diepvens F, Pals G (2002) Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. Nucleic Acids Res 30:e57CrossRefPubMedGoogle Scholar
  2. 2.
    2. van Dijk MC, Rombout PD, Boots-Sprenger SH, Straatman H, Bernsen MR, Ruiter DJ, Jeuken, JW (2005) Multiplex ligation-dependent probe amplification for the detection of chromosomal gains and losses in formalin-fixed tissue. Diagn Mol Pathol 14:9–16CrossRefPubMedGoogle Scholar
  3. 3.
    3. Takata M, Suzuki T, Ansai S, Kimura T, Shirasaki F, Hatta N, Saida T (2005) Genome profiling of melanocytic tumors using multiplex ligation-dependent probe amplification (MLPA): Its usefulness as an adjunctive diagnostic tool for melanocytic tumors. J Dermatol Sci 40:51–57CrossRefPubMedGoogle Scholar
  4. 4.
  5. 5.
    5. Hopman AH, van Hooren E, van de Kaa CA, Vooijs PG., Ramaekers FC (1991) Detection of numerical chromosome aberrations using in situ hybridization in paraffin sections of routinely processed bladder cancers. Mod Pathol 4:503–513PubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Minoru Takata
    • 1
  1. 1.Shinshu University School of MedicineMatsumotoJapan

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