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A phenol technique for extraction of alkylated DNA, RNA, and protein from a single tissue sample

✍ Scribed by Lawrence C. Panasci; Dianna C. Green; Patricia A. Fox; Philip S. Schein

Publisher
Elsevier Science
Year
1977
Tongue
English
Weight
594 KB
Volume
83
Category
Article
ISSN
0003-2697

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✦ Synopsis

A procedure for extracting DNA, RNA, and protein from an L1210 leukemia cell sample is described. Sodium deoxycholate and 154 mM NaCl in the presence of phenol extracts cytoplasmic RNA. Sodium deoxycholate, paminosalicylate, and 1 M NaCl in the presence of phenol extracts the DNA. The DNA is then freed of RNA with RNase Tl and T2. Cytoplasmic RNA with no detectable DNA and less than 0.02% protein contamination was so obtained. The DNA isolated had less than 3% uracil as compared to thymine and no more than 0.1% protein. The extraction procedures are especially useful for investigating nucleotide alterations in nucleic acids by chemical carcinogens and alkylating agents in tissue samples of only 2 x IO8 cells. The procedure has also been successfully applied to murine bone marrow cells, human bone marrow cells, HeLa cells, and myeloblasts from a patient with acute myelogenous leukemia.

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