using ficoll to collect mononuclear lymphocytes - IS it suitable for DNA extraction?? (Apr/16/2008 )

i want to know if there any problem in separation of lymphocytes using ficoll before extraction of DNA ready for PCR method. ????

and if i collect lyphocytes and wash it by PBS
how can i preserve it for a long time???? and could i keep it at -20 or -70 C???

Best regards.

is there is no problem to use Ficoll????


and i want to know how to keep lymphocytes after separation for a longer time before doing DNA extraction???

Ficoll usually is used to create heavier density for DNA isnt it?

I thought ficoll is used for precipitate the nucleic acid? no? Perhaps I am wrong.

sorry then.. i mistaken it with some other name. cant help u then. good luck!

If used carefully then ficoll will seperate your blood no problem. If you carefully layer your blood on top of the ficoll and then centrifuge, you should get Monocytes in the middle of the ficoll and PMN's/ Erythrocytes (RBC's) pelleted at the bottom of the tube. You can then lyse the RBC's with Ammonium Chloride.

By carefully I mean very very very slowly add the blood to the ficoll. If you add it too quickly the whole blood will contaminate the lower part of the non centrifuged ficoll, and will result in an unpure separation.

Hope this is useful.

Rhombus