Article

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

Medizinische Hochschule Hannover, Institute of Pathology, Hannover, Germany.
Methods in molecular biology (Clifton, N.J.) 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.

0 0
 · 
0 Bookmarks
 · 
64 Views
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.

Keywords

convenient storage
 
efficient mRNA retrieval
 
isolating anatomic compartments
 
mRNA extraction
 
mRNA quality
 
paraffin-embedded tissues
 
PCR)-based expression analysis
 
significant loss
 
stable preservation
 
staining
 
subsequent RT-polymerase chain reaction
 
tissue samples
 
well-established