Extraction of RNA from single frozen sections.
The sensitivity of the reverse transcriptase-polymerase chain reaction (RT-PCR) makes it ideally suited for the detection of changes in gene expression. Unfortunately, traditional methods for RNA isolation require time-consuming procedures that are not appropriate for small samples, such as individual frozen sections. This report describes a new technique that permits the rapid extraction of RNA from individual frozen histological sections. RNA is extracted by incubating a frozen section in an RT-PCR compatible buffer solution containing RNase inhibitor and dithiothreitol. RNA isolated from frozen sections is stable at room temperature for up to 3 h under the conditions described. Alternatively, extracts can be frozen for later use. When maintained in a dry state at room temperature, RNA in sections remained stable for 2 weeks. Histological, immunohistochemical, or in situ analyses can be carried out with sections that are immediately adjacent to those used for extracting RNA. The simplicity and economy of this procedure may foster the development of prognostic screens that can be performed in parallel with traditional histopathological analysis.
Taylor
G P
GP
Department of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio 78284, USA.
Troyer
D A
DA
Giambernardi
T A
TA
Klebe
R J
RJ
D002021:Buffers; D015925:Cryopreservation; D004229:Dithiothreitol; D004791:Enzyme Inhibitors; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D011467:Prostate / Q000737:chemistry*; D012333:RNA, Messenger / Q000302:isolation & purification*; D012260:Ribonucleases / Q000037:antagonists & inhibitors; D012996:Solutions; D013997:Time Factors
D002021:Buffers; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D012996:Solutions; D012260:Ribonucleases; D004229:Dithiothreitol