Proper storage of isolated white blood cell prior to RNA isolation
Posted 12 October 2011 - 04:07 AM
I am new in the field of PCR and all the subprocedures so I hope that someone can help me.
Accoring to my plan I am going to isolate peripheral blood leukocytes from whole rat blood by the use of histopague 1077 methods. I need to store the sample proper before RNA isolation. The question is can I store the sample in the freezer (-80) for a couple of mount or do I need to use RNAlater?
Second; Do I need do something about the RNases?
Best regards Sophie
Posted 12 October 2011 - 04:29 AM
I used to just add Trizol to my cells and then put in the -80C until extraction. Never had a problem with that, and I've never used RNAlater.
Hope this helps.
Posted 13 October 2011 - 01:29 AM
Yes, I am going to extract RNA.
The first step will be the leukocytes isolation from whole blood. The second step, RNA isolation, will be performed in the beginnig of 2012.
Further, there have been some discussion about the use of sterile equipment for the leukocyte isolation. If I take following precautions which include the use of gloves and mask do I need to buy sterile, centrifuge tubes, pipetts ect?