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

Authors :
Jonigk D
Modde F
Bockmeyer CL
Becker JU
Lehmann U
Source :
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2011; Vol. 755, pp. 67-75.
Publication Year :
2011

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.

Details

Language :
English
ISSN :
1940-6029
Volume :
755
Database :
MEDLINE
Journal :
Methods in molecular biology (Clifton, N.J.)
Publication Type :
Academic Journal
Accession number :
21761294
Full Text :
https://doi.org/10.1007/978-1-61779-163-5_5