2 replies to this topic

[felixwang]

I am planning to do doing stable transfection of Drosophila Kc-167 cells, using pMK33 vector. Is there anyone here has some experience of the procedure? I got a protocol which recomended that after 24 hr, splited the cells 1:4 into new medium and after another 24 hr, add the selective drug. I am wondering why we have to split cells 24 hr after transfection? Why not wait until cells are confluent?

[fred_33]

hi
generally, cells need to be active to be selected with an antibiotic. that's why they recommend to divide them the day after transfection. But if you have enough place on your plate for good division, you can do without split.

[felixwang]

Thanks Fred!