256 



G. R. WYATT 



the Rf value of yeast adenylic acid is not recorded. Adenosine-5'-phosphate, adeno- 

 sine diphosphate, and adenosine triphosphate are also separable from one another in 

 this system, or, with rather better spacing, in a mixture of 65 vol. of acetone with 35 

 vol. of 15% trichloroacetic acid. 



From the published Rp values, the buffered isoamyl alcohol-tetrahydrofurfuryl 



TABLE II 



Rf Values of Nucleotides 



" tert-B\ita.no\ at 26° 700 ml., const.-boiling HCl 132 ml., water to make 1 liter; Whatman No. 1, descend- 

 ing; Smith and Markham, Biochem. J. '46, 509 (1950). Rf values (20-22°) are from Boulanger and Montreuil, 

 Bull. soc. chim. biol. 33, 784, 791 (1951). 



'' See footnote /, Table I. The value for orthophosphate is from Markham and Smith, Biochem. J. 49, 

 401 (1951). 



' Acetone 75 vol., 25% (wt./vol.) trichloroacetic acid 25 vol.; Whatman No. 1 paper washed in 2 A'' HCl 

 and water, ascending, at 4°; Burrows et al., Nature 170, 800 (1952). 



"* Isoamyl ale. 1 vol., tetrahydrofurfuryl ale. 1 vol., 0.08 M potassium citrate buffer (pH 3.02) 1 vol.; What- 

 man No. 1, descending, 20-25°; Carpenter, Anal. Chem. 24, 1203 (1952). 



* Isobutyric acid 10 vol., 0.5 A^ NH4OH 6 vol., pH 3.6-3.7; Schleicher and Schiill No. 597, descending, 

 21-25°; Magasanik et al., J. Biol. Chem. 186, 37 (1950). Rp values calcd. from the published relative mobilities 

 and figures. 



^ 90% aq. phenol 84 vol., <er<-butanol 6 vol., formic acid 10 vol., water 100 vol.: after sepn. at a temp. 

 2-3° below that of the chromatography room, the nonaq. phase is used and the aq. phase is placed in the bot- 

 tom of the tank; Whatman No. 1 paper, descending, 20-22°; Boulanger and Montreuil, op. cit. 



" See footnote i. Table I. Values for adenosine-5'-phosphate, diphosphate, and triphosphate are from 

 W.E. Cohn and C. E. Carter, J. Am. Chtm. Soc. 72, 4273 (1950). 



'' Satd. (NHOiSOi in water 79 vol., 0.1 M buffer soln. (pH 6) 19 vol., isopropanol 2 vol.; Whatman No. 1 

 paper, descending; Markham and Smith, op. cit. Rp values are measured from the published figure. 



* The "a" and "b" nucleotides of Carter, J. Am. Chem. Soc. 72, 1466 (1950), and subsequent authors are 

 now identified as the 2'-phosphates and 3'-phosphates, respectively, of the ribosides. 



