Optimized RNA extraction from non-deparaffinized, laser-microdissected material.

Medizinische Hochschule Hannover, Institute of Pathology, Hannover, Germany.
Methods in molecular biology (Clifton, N.J.) (Impact Factor: 1.29). 01/2011; 755:67-75. DOI: 10.1007/978-1-61779-163-5_5
Source: PubMed

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.

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