Chromatographic separation of mononucleotides derived from transfer ribonucleic acids.
1. A method is described for fractionating alkaline and enzymic hydrolysates of ribonucleic acids and oligonucleotides on DEAE-cellulose columns by using a constant-composition eluent. 2. The relative partition coefficients of the four major and several minor nucleotides present in nucleic acid digests are given. 3. The influence of pH and of molarity of the buffer on the separation is described. 4. Relations for the quantitative determination of the major nucleotides from their absorption zone areas on the chromatographic record are derived. 5. Hydrolysis of oligonucleotides with T2 ribonuclease yields solely the nucleoside 3'-monophosphates, thereby simplifying the subsequent chromatographic separation. 6. Examples of the qualitative and quantitative analysis of oligonucleotides from enzymic hydrolysates of RNA are given.
D002848:Chromatography, DEAE-Cellulose; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008722:Methods; D008970:Molecular Weight; D009711:Nucleotides / Q000302:isolation & purification*; D011119:Polynucleotides / Q000302:isolation & purification; D012313:RNA / Q000302:isolation & purification; D012343:RNA, Transfer / Q000032:analysis*; D012260:Ribonucleases
D009711:Nucleotides; D011119:Polynucleotides; D012313:RNA; D012343:RNA, Transfer; D012260:Ribonucleases