3 replies to this topic

[Yasar]

Hi,
I am looking at BCR/ABL expression of two philadelphia positive cell lines. According to plan, I have to use three clones of each cell line and do a PCR in triplicates. How long do I have to grow a cell line in three different flasks to call them different clones of a cell line? Is there any guideline or definition
Thanks
Yasar

[Kamran]

Three clones of a single cell means, you pick up single cell from each cell line, grow those single cells to a desired cell density and perform your experiment.

[Yasar]

Hi Kamran,
Thank you very much for your reply. how would I pick one cell from an aliquot of cells and how would I grow them? Is there any protocol for that? I thawed three frozen vials of a cell lines and grew them separately. Would you consider them as three clones?
Thanks.

[bob1]

No, three vials are not considered clones.  Have a look for protocols relating to limited dilution and/or colony selection.