Isolation of messenger RNA from polysomes by chromatography on oligo(dT)-cellulose

✍ Scribed by R.E. Pemberton; P. Liberti; C. Baglioni

• Publisher: Elsevier Science
• Year: 1975
• Tongue: English
• Weight: 665 KB
• Volume: 66
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(75)90720-4

✦ Synopsis

A method for the isolation of messenger RNA (mRNA) from polysomes is described. Pofysomes are dissolved in a solution containing 0.5 M NaCl and Na dodecyl sulphate and applied to an oligo(dT)-cellulose column. RNA species containing poly(A) sequences are retained by the column, whereas ribosomal proteins and other RNA species are washed off. The column is then eluted with a buffer not containing NaCI. mRNA from HeLa cells and from duck reticulocytes has been fractionated in this way. When fractionated on sucrose gradients, 10 s globin mRNA is obtained in addition to a 20 s component, which can be translated in a cell free system into duck globin. This 20 s RNA is an aggregate of mRNA, which can be disaggregated. Experiments with HeLa cells have shown that the only mRNA species which is not retained by oligo(dT)-cellulose is histone mRNA; this mRNA does not contain a poly(A) sequence. 18