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Optimized RNA extraction from non-deparaffinized, laser-microdissected material.
- 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.
- Subjects :
- Cell Separation methods
Data Interpretation, Statistical
Gene Expression
Gene Expression Profiling methods
Genes, Wilms Tumor
Humans
Kidney Glomerulus cytology
Kidney Glomerulus metabolism
Membrane Proteins genetics
Membrane Proteins metabolism
Microtomy methods
Pulmonary Veins cytology
RNA Polymerase II genetics
RNA Polymerase II metabolism
RNA, Messenger genetics
Reverse Transcriptase Polymerase Chain Reaction methods
Tissue Fixation methods
Umbilical Arteries cytology
Lasers
Microdissection methods
Paraffin Embedding
RNA, Messenger isolation & purification
Subjects
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