Separation of 5′-ribonucleotides by two-dimensional thin-layer chromatography

✍ Scribed by R.G. Stickland

• Publisher: Elsevier Science
• Year: 1965
• Tongue: English
• Weight: 656 KB
• Volume: 10
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(65)90244-7

✦ Synopsis

Recently simple mixtures of two nucleotides were separated in as little as 10 min by one-dimensional thin-layer chromatography, but in most cases complex mixtures of more than about four nucleotides have not been resolved (l-3). With paper or column chromatography all the 5'-mono-, di-, and triphosphates of adenosine, cytidine, guanosine, and uridine can be separated but this usually takes many hours.

A method is described in this paper whereby these twelve nucleotides can be separated into eleven spots by two-dimensional thin-layer chromatography in 3.5 to 4 hr. CTPl and UDP are not resolved. Polysaccharide and fluorescent material (as found in biological samples, for example rat liver homogenate) were found to interfere with this chromatography and it was first necessary to remove these substances by chromatography on thin-layers of Dowex-1 plus Sephadex G-25.

MATERIALS AND METHODS

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