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RNA won't dissolve in DEPC water - (Sep/27/2006 )

Hai,

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.

Thank you
Rukmini

-radhu-

I incubate the RNA at 55-60 degrees C for 10 min to dissolve the pellet completely.

-ucsdgrad88-

also, perhaps the pH of your water is too low?

-aimikins-

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?

Thank you
Rukmini

-radhu-

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.

-fred_33-