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Optimized RNA Extraction from Non-deparaffinized, Laser-Microdissected Material

  • Danny Jonigk
  • Friedrich Modde
  • Clemens L. Bockmeyer
  • Jan Ulrich Becker
  • Ulrich LehmannEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 755)

Abstract

mRNA extraction and subsequent RT-polymerase chain reaction (PCR)-based expression analysis from laser-microdissected material is by now a well-established and reproducible method. Most routinely stored tissue samples are preserved as formalin-fixed, paraffin-embedded materials. While this allows for a convenient storage and stable preservation of nucleic acids, deparaffinization before staining for laser microdissection may result in a significant loss of mRNA quality and consequently of PCR sensitivity. We describe a method of isolating anatomic compartments from non-deparaffinized, formalin-fixed, and paraffin-embedded tissues by laser-assisted microdissection which allows for a highly efficient mRNA retrieval.

Key words

mRNA extraction Non-deparaffinized sections Laser-assisted microdissection Quantitative RT-PCR 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Danny Jonigk
    • 1
  • Friedrich Modde
    • 1
  • Clemens L. Bockmeyer
    • 1
  • Jan Ulrich Becker
    • 1
  • Ulrich Lehmann
    • 1
    Email author
  1. 1.Medizinische Hochschule HannoverInstitute of PathologyHannoverGermany

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