Step 3 Purify RNA on a Qiagen spin column

1) Transfer the clear aqueous phase into a new RNase-free tube. Add 0.5 ml isopropyl alcohol per 600 ^l of isolated aqueous phase. The correct final alcohol concentration is critical for maximal binding to the column matrix.

2) Place solution on a Qiagen RNeasy mini or midi column (750 ^l at a time). Spin 30 sec at 13,000 RPM reloading with remaining solution until all has passed through the column.

3) Continue with the washing and elution of the RNA as outlined in the Qiagen instructions.

4) Elute the final RNA in DEPC-H2O.

Note: other column types can be used but the alcohol concentration has to be compatible, otherwise the RNA may not stick to the column matrix.

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