RNA Purification - After DNase (Sep/02/2009 )
Anyone know if i can use Qiagen's qiaquick purification to purify ssRNA?
I need to remove the EDTA from my DNasing step
will phenol purification also work?
You can use any kit for RNA purification, but generally for obtaining high quality RNA without loosing much RNA, we use LiCl (lithium chloride) method to clean the RNA. So let me give the procedure:
1 volume of 7,5 M lithium chloride
1 volume of DNAse treated RNA sample
1 volume of nuclease free water
(so final concentration of lithium chloride will be 2,5 molar)
mix them, put -20 for at least 1 hour, overnight is always better, centrifuge at 15,300 rpm (or max speed on microcentrifuge) for 15-20 min at +4 C, discard the supernetant, add 1 ml of 70% EtoH (it should be -20 chilled), centrifuge at max speed for 15-20 min., discard the supernatant, dry the pellet, resuspend with nuclease free water.
it seems a bit long but to be in safe side this is the best way
that s all
Thanks a lot!!! Glad someone could help me out here.