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RNAqueous-96 RNA Isolation Kit Automation Protocol


This protocol was developed for the MultiPROBE II HT Liquid Handling System with a Gripper Integration Platform. Time required: 1 hr 20 min.

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)
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