Isolation of poly(A)-containing RNA on synthetic fluorophlogopite (mica)

✍ Scribed by Peter Pulkrabek; Kamil Klier; Dezider Grunberger

• Publisher: Elsevier Science
• Year: 1975
• Tongue: English
• Weight: 519 KB
• Volume: 68
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(75)90675-2

✦ Synopsis

Synthetic fluorophlogopite, an aluminosilicate of the same structure as naturally occurring mineral mica in which potassium ions on the basal surface have been replaced by aluminum ions, has the ability to retain polynucleotides irreversibly. This property of Al:J+-mica was used for irreversible adsorption of poly(U) and subsequent selective adsorption of poly(A)-containing RNA from rabbit reticulocyte polysomes at high salt concentration and its elution by 50% dimethylsutfoxide. The properties of RNA isolated on poly(U)-AP-mica were studied by sucrose density gradient centrifugation and by stimulation of globin synthesis in an in vitro protein synthesizing system from wheat germ and from Krebs II-ascites cells. The preparation contained 9s RNA species which corresponds to rabbit globin messenger RNA, and under optimal conditions it stimulated protein synthesis more than 100-fold. Polyacrylamide-gel electrophoresis in sodium dodecyl sulfate showed that synthesized product was identical with rabbit globin.