Purification of polyA RNA by the Oligotex
Material
D Sol.
10% suspension of Oligotex: 50μl suspension / 500 μg total RNA
Wash buffer: 10 mM Tris-HCl (pH7.5), 150 mM NaCl, 1 mM EDTA
5M NaCl
Elution Buffer: 5 mM Tris-HCl
Glycogen solution
Method
□ Dissolve total RNA in D solution in 2 ml eppendorf tube
□ Add more than x2 volume of elution buffer
□ Add Oligotex suspension: 50μl suspension / 500 μg total RNA
□ Heat at 70°C for 5 min
□ Add 1/10 volume of 5M NaCl
□ Incubate at 37°C for 5 min and centrifuge
□ Wash with 1 ml of wash buffer once.
□ Add 400 μl of Elution buffer and incubate at 70°C for 5 min and centrifuge (Max for 20 min).
□ Recover the sup.
□ Centrifuge in spin column to eliminate the contaminant oligotex
□ Add 1 μl of glycogen, 1/10 volume of Na+-Ac, and 3 volume of EtOH
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