Hello Menda Rajendar
Cumulus cells should be washed with PBS after collection, and then you may store these cells at −80°C until RNA extraction.
Best.
Thanks for suggestion but is any storage media for this? and after collection can I store liquid nitrogen ?
If you are worried about RNA, take the cumulus without dissociating. Pick up with fine pasteur pipette or pipettor 20-40ul tip (better) Give a rinse in RNALater in a small dish. And pool all the cumuli in 1ml of RNALater in a sterile tube. Follow instructions for storage on RNAlater sheet (enclosed). I will recommend not to dissociate cumulus and not to wash. More manipulations will change your gene expression. After thawing, spin , get rid of RNALater and go with Trizol directly and follow RNA extraction method or any other method you are trying to follow. I have used RNAlater many times, that is very useful.
Good luck
if you have loose cumulus cells, than spin and discard the supernatant, than give one wash with RNALAter, rest same as above
I am glad to to you sir, its very difficult to collect the cumulus cells with out dissociation. while vigor's pipetting automatically cumulus cells will dissociate and i will store only loose cumulus cells only but after centrifugation pellet will get at bottom of the tube and how to store the pellet ? is should i store in RNALAter sir ?
To the pellet of cells, add 1ml of RNALater and store -80C or LN2, you can store for years, Good luck
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