Extraction of RNA from single frozen sections - PubMed (original) (raw)
Extraction of RNA from single frozen sections
G P Taylor et al. J Pathol. 1998 Mar.
Abstract
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.
Similar articles
- [Use of the real-time RT-PCR method for investigation of small stable RNA expression level in human epidermoid carcinoma cells A431].
Nikitina TV, Nazarova NIu, Tishchenko LI, Tuohimaa P, Sedova VM. Nikitina TV, et al. Tsitologiia. 2003;45(4):392-402. Tsitologiia. 2003. PMID: 14520871 Russian. - [Quick RNA extraction from specified morphological region in frozen sections and its RT-PCR application].
Jang Y, Liu N, Liu J, Li H. Jang Y, et al. Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2000 Feb;17(1):47-50. Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2000. PMID: 10653910 Chinese. - A simple method for fixation and microdissection of frozen fresh tissue sections for molecular cytogenetic analysis of cancers.
Huang Q, Sacks PG, Mo J, McCormick SA, Iacob CE, Guo L, Schaefer S, Schantz SP. Huang Q, et al. Biotech Histochem. 2005 May-Aug;80(3-4):147-56. doi: 10.1080/10520290500382806. Biotech Histochem. 2005. PMID: 16298900 - RT-PCR without RNA isolation.
Klebe RJ, Grant GM, Grant AM, Garcia MA, Giambernardi TA, Taylor GP. Klebe RJ, et al. Biotechniques. 1996 Dec;21(6):1094-100. doi: 10.2144/96216rr02. Biotechniques. 1996. PMID: 8969838 - Assessment of gene expression in head and neck carcinoma using laser capture microdissection and real-time reverse transcription polymerase chain reaction.
Malhotra PS, Malekfzali A, Bonner RF, Juhn S, Van Waes C, Chen Z. Malhotra PS, et al. Laryngoscope. 2004 Dec;114(12):2123-8. doi: 10.1097/01.mlg.0000149446.14770.52. Laryngoscope. 2004. PMID: 15564832
Cited by
- Immuno-LCM: laser capture microdissection of immunostained frozen sections for mRNA analysis.
Fend F, Emmert-Buck MR, Chuaqui R, Cole K, Lee J, Liotta LA, Raffeld M. Fend F, et al. Am J Pathol. 1999 Jan;154(1):61-6. doi: 10.1016/S0002-9440(10)65251-0. Am J Pathol. 1999. PMID: 9916919 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources