Procedure for the base composition of RNA especially suited for samples containing relatively large amounts of DNA and protein
✍ Scribed by Charles W. Lees
- Publisher
- Elsevier Science
- Year
- 1971
- Tongue
- English
- Weight
- 654 KB
- Volume
- 40
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
Biological samples containing large amounts of DNA relative to RNA present considerable difEculty to the analyst who wishes to determine the base composition of the RNA. Direct analysis is obviously ruled out since the bases, A, G, and C2 are common to both molecules. Several general approaches to the separation and determination of nucleic acids have been reviewed (1). Mild alkaline hydrolysis (2)) which separates RNA from the bulk of the DNA and protein, is especially applicable to the circumstance of large amounts of DNA. In this paper, several modifications of this step were tested and the one giving the lowest DNA background was selected to precede acid hydrolysis, ion-exchange separation, and spectrophotometric determination.
The combined procedure is inexpensive, accurate, and relatively rapid. MATERIALS Chemicals were reagent grade unless otherwise specified. The water was house-distilled and then redistilled in an all-glass still and stored in a glass reservoir. Adenine, G, CMP (2'3 mixed), UMP (2'-3' mixed), CR, UR, DNA (calf thymus, type I), RNA (yeast, type XI), and bovine serum albumin were purchased from Sigma Chemical Company. Yeast RNA (7GA) was purchased from the Worthington Biochemical Corporation. All absorbance measurements were carried out using a Zeiss PMQII spectrophotometer. The pH was measured with a Radiometer 28 pH meter with a combination electrode. Thin-layer chromatography was performed ' Supported