RNAqueous(tm)-96 RNA Isolation Kit Automation Protocol - Biocompare Buyer's Guide For Life Scientists

RNAqueous™-96 RNA Isolation Kit Automation Protocol

This protocol was developed for the MultiPROBEŒ 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).

Add 150 µl Lysis Solution to each well.
Add 150 µl 64% ethanol to each well.
Mix and transfer to the 96-well filter plate.
Apply vacuum suction (20 mm Hg) for 3 min (RNA remains bound to the filter).
Add 400 µl Wash Solution A to each well and apply vacuum suction (20 mm Hg) for 2 min.
Add 40 µl (100 units) DNase I.
Incubate 20 min at room temperature.
Add 400 µl Wash Solution B to each well and apply vacuum suction (20 mm Hg) for 2 min.
Add 400 µl Wash Solution C/D to each well and apply vacuum suction (20 mm Hg) for 2 min.
Add 400 µl 100% ethanol to each well and apply vacuum suction (20 mm Hg) for 2 min.
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).
Move Spacer and Collection plates to the vacuum manifold. Place 96-well filter plate on top.
Add 80 µl Elution Buffer to each well.
Apply vacuum suction at 10 mm Hg for 30 sec then at 20 mm Hg for 2 min (RNA is transferred to collection plate).
(optional) Manually check for fluid remaining on filter plate. If fluid is present, centrifuge the filter plate/collection assembly to recover residual sample.