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a friend of mine had the same problem last time with heat inactivation. above 58C he got solid phase. then I think he increased temp from 56 to 57 in hot block (not water bath) and then tapped every 5-10 min to make sure the serum mixed.
#154245 How to test heat inactivated Normal Human Serum
Posted
on 25 April 2013 - 11:03 PM
#138581 Understanding Cell Passaging
Posted
on 30 July 2012 - 12:48 PM
CMIRC, on 30 July 2012 - 06:22 AM, said:
If I thaw a vial of PC3 cells (prostate cancer immortal cell line) and culture them. The passage number would be 0 when I thaw them first. However the passage number would increase to 1 when I sub culture them. Say after subsequent cell culturing I decide to freeze the cells at passage 4. The next time I take out a the PC3 vial from liquid nitrogen and culture would the passage start from 4 or from 0?
CMIRC, on 30 July 2012 - 06:22 AM, said:
I know that for primary cell lines the passage number does not reset and continous even after freezing and thawing. however does the same apply for immortal cancer cells. And if it does then upto what passage can one do experiments without having any genetic alterations due to the passaging. ie upto what passage can one use an immortal cell line before a buying a new one?
CMIRC, on 30 July 2012 - 06:22 AM, said:
Also for non adherent cell lines when would a passage be considered. If I take flask of 50% confluent THP1 cells and centrifuge them and put them back in the same flask with new media without splitting would that be a passage, secondly if I have 2 flasks of THP1 cells 25% confluence and I put them into one flask without changing the medium or centrifuging, then would that be a passage?
