Purification of poly-A RNA

In the following a slightly modified version of the conventional oligo(dT)-cellulose chromatography procedure suitable for obtaining poly-A+ RNA from about 1-10 mg total RNA is presented. 

2% SDS Medium-salt buffer 10 mM Tris-HCl, pH 7.5 100 mM NaCl 1 mM EDTA 0.1% SDS Elution buffer 10 mM Tris-HCl, pH 7.5 1 mM EDTA 0.1% SDS Wash buffer 

Suspend 0.2 g oligo(dT)-cellulose powder in 2 ml wash buffer and pour into the column.3 

Rinse the packed column with 15 ml H20 or until pH of the effluent is 8.5. 

Collect the flow-through, leave it at 65°C for 5 min and reapply to the column. 

---