RNA won't dissolve in DEPC water - (Sep/27/2006 )
I have been trying to do total RNA isolation, in the final step that is while dissolving the pellet in DEPC treated water, a transparent jelly like remains back. I have isolated it for Jurkat JR4 cells. I have washed the final with 75% ice cold ethanol
could please suggest a way to dissolve the whole pellet in DEPC treated water.
I incubate the RNA at 55-60 degrees C for 10 min to dissolve the pellet completely.
also, perhaps the pH of your water is too low?
Thank y ou very much for the reply, can iplease know what is the pH range and the buffer in which RNA pellet gets dissolved?
normally such white gel appears when the RNA was too dried.
So incubation in DEPC treated water should be done longer in ice, for rehydrating the RNA pellet. Then 60° 10' should end the job.
pH range of your water should be 7. Aimee means that it's often that pH from miliQ water goes down to 5 due to air exposure.
but normally should affect the resuspension.