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KMID : 0364519960080010037
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Dong-A Journal Medicine
1996 Volume.8 No. 1 p.37 ~ p.44
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Study on Extraction of Nucleolar RNA and snRNA
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Abstract
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There are two popular RNA extraction methods, presently employed by a member of investigators. One is the hot SDS/phenol method which requires the use of 0.3% SDS for RNA extraction at pH 5.1, followed by the use of phenol for deproteinization at
high
temperature of 63℃. The other is the use of RNAsol(guanidine thiocyanate/phenol) both for RNA extraction and deproteinization. The latter is known to be very useful for mRNA preparation. These two methods were investigated to characterize the
types of
RNA extracted from isolated HeLa cell nuclei, according to the sizings of RNA species by 1.5% agarose gel electrophoresis, 5-40% sucrose density gradient centrifugation and 10% polyacylamide gel electrophoresis. It was observed that the RNA
species
extracted by the RNAsol method showed lower efficacy in the extraction of nucleolar high molecular weight RNAs and snRNAs. Using the hot SDS/phenol method, the nucleolar 45S pre-rRNA and snRNAs were efficiently extracted.
Accordingly, it is recommended that the hot SDS/phenol method is to be used for the study of snRNAs and nucleolar high molecular weight RNAs.
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KEYWORD
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