Abstract：
:Microarray technologies have made possible comprehensive analyses of nucleic acid sequence and expression. However, the technology to obtain efficiently high-quality RNA and DNA suitable for array analysis from purified populations of neoplastic cells from human tissues has not been well addressed. Microdissection can enrich for populations of cells present in various tumor tissues, but it is not easily automated or performed rapidly, and there are tissues in which cells of interest cannot be readily isolated based on morphologic criteria alone. Here we describe a protocol for efficient RNA and DNA isolation from flow cytometrically purified whole epithelial cells from primary tissue. The aqueous reagent, RNAlater, which preserves RNA, allows immunolabeling and purification of whole epithelial cells by flow sorting without special instrument preparation to reduce RNase activity. We used real-time PCR to determine RNA quality afterflow sorting. High-quality RNA and DNA suitable for expression and genotype analysis can be readily obtained from flow cytometrically purified populations of neoplastic cells from human tissues.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Barrett MT,Glogovac J,Prevo LJ,Reid BJ,Porter P,Rabinovitch PSdoi
10.2144/02324rr06subject
Has Abstractpub_date
2002-04-01 00:00:00pages
888-90, 892, 894, 896issue
4eissn
0736-6205issn
1940-9818journal_volume
32pub_type
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