( This protocol was developed f...)RNAqueous-96 RNA Isolation Kit Automation Protocol

For more information on the protocol, see the RNAqueous-96 Instruction Manual (pdf).

1. Add 150 l Lysis Solution to each well.

2. Add 150 l 64% ethanol to each well.

3. Mix and transfer to the 96-well filter plate.

4. Apply vacuum suction (20 mm Hg) for 3 min (RNA remains bound to the filter).

5. Add 400 l Wash Solution A to each well and apply vacuum suction (20 mm Hg) for 2 min.

6. Add 40 l (100 units) DNase I.

7. Incubate 20 min at room temperature.

8. Add 400 l Wash Solution B to each well and apply vacuum suction (20 mm Hg) for 2 min.

9. Add 400 l Wash Solution C/D to each well and apply vacuum suction (20 mm Hg) for 2 min.

10. Add 400 l 100% ethanol to each well and apply vacuum suction (20 mm Hg) for 2 min.

11. Move 96-well filter plate to dry heat block covered with fresh paper towels and heat at 42C for 3 min (this removes any residual wash solution).

12. Move Spacer and Collection plates to the vacuum manifold. Place 96-well filter plate on top.

13. Add 80 l Elution Buffer to each well.

14. Apply vacuum suction at 10 mm Hg for 30 sec then at 20 mm Hg for 2 min (RNA is transferred to collection plate).

15. (optional)
    '"/>