title: Optimized RNA extraction from non-deparaffinized, laser-microdissected material.
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.
authors.affiliation: Medizinische Hochschule Hannover, Institute of Pathology, Hannover, Germany.
authors.forename: Danny
authors.initials: D
authors.lastname: Jonigk
authors.forename: Friedrich
authors.initials: F
authors.lastname: Modde
authors.forename: Clemens L
authors.initials: CL
authors.lastname: Bockmeyer
authors.forename: Jan Ulrich
authors.initials: JU
authors.lastname: Becker
authors.forename: Ulrich
authors.initials: U
authors.lastname: Lehmann
mesh_terms: D002469:Cell Separation / Q000379:methods; D003627:Data Interpretation, Statistical; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods; D016162:Genes, Wilms Tumor; D006801:Humans; D007678:Kidney Glomerulus / Q000166:cytology / Q000378:metabolism; D007834:Lasers; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D042282:Microdissection / Q000379:methods*; D008867:Microtomy / Q000379:methods; D016612:Paraffin Embedding; D011667:Pulmonary Veins / Q000166:cytology; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D016707:Tissue Fixation / Q000379:methods; D014469:Umbilical Arteries / Q000166:cytology
chemical_list: D008565:Membrane Proteins; D012333:RNA, Messenger; C113267:nephrin; D012319:RNA Polymerase II