how can i preserve lymphocytes after isolation - keep lymphocyte pellets at -70 C ? (Apr/30/2008 )

PLZ i want to know how to keep lymphocytes pellet after its isloation with Histopaque technique.

can i keep the pellets at -70 C for furthur DNA extraction???

or add PBS to the Pellets

or what else????

Best regards

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If you just need to prepare nucleic acid from the cells, wash the cells after isolation with PBS, remove the supernatant after the spin and store the pellet at -80˚C.

You can also resuspend the pellet in RNAlater if it is crucial you preserve the transcripts. Store at 4˚C for up to a week or transfer to -80˚C after being overnight at 4˚C.

Hope this helps,

AussieUSA.

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thank you so much for your reply. i have isloted lymphocytes then wash by PBS then add RBC's lysis buffer to get rid of excess RBCs then discard supernatant and preserve lymphocytes' pellets at -70 C. is it right or should i wash by PBS before preservation????



sorry i don't understand what do you mean by this words.

Best regards.

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i have isloted lymphocytes then wash by PBS then add RBC's lysis buffer to get rid of excess RBCs then discard supernatant and preserve lymphocytes' pellets at -70 C. is it right or should i wash by PBS before preservation????

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I have isolated lymphocytes and my further downstream application was isolation of RNA, so i was advised to store the lymphocytes in trizol solution.

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thank you so much for your kind reply.
But i need to do DNA extraction so how can i keep lymphocytes for furthur extraction of DNA?????

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i have isloted lymphocytes then wash by PBS then add RBC's lysis buffer to get rid of excess RBCs then discard supernatant and preserve lymphocytes' pellets at -70 C. is it right or should i wash by PBS before preservation????

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