RNA cleanup with magnetic beads
Dominik Buchner
Abstract
This protocol describes cleaning up RNA extracts with carboxylated magnetic beads and a PEG-NaCl buffer. It can also be used for volume reduction of a sample or buffer exchange after enzymatic reactions (e.g. DNAse treatment).
Before start
Make sure all buffers are prepared before starting.
For more effortless pipetting let the bead solution adjust to Room temperature
Steps
Shake the RNA cleanup solution until the beads are homogeneously resuspended
To 100µL add 200µL in a 1.2 mL Deep Well Storage Plate
To bind the RNA to the beads shake at 900rpm
Place the plate on a magnet to pellet the beads for 0h 5m 0s or until the mixture appears clear
Discard the supernatant by pipetting
With the plate still on the magnet, add 100µL to each sample
Incubate for at least 0h 0m 30s
Discard the supernatant by pipetting
With the plate still on the magnet, incubate the plate for 0h 5m 0s at Room temperature to dry off residuals of wash buffer
Add 100µL to each sample
900rpm to elute the RNA from the beads
Place the plate on a magnet to pellet the beads for 0h 5m 0s
Transfer 100µL of the eluted RNA to a new storage plate. Store at -80°C
