making stable clones through single cell dilution after transfection - (Jul/28/2006 )

I am trying to make stable clones of transfected CHO cells thru single cell dilutions and I was wondering how long after putting the cells into high conc. G418 do people normally do thier single cell dilution?

Also, I am not too familiar with single cell dilutions so i wanted to know if anyone had any suggestions and tips from past experience...i was planning on diluting my transfected cells to 0.5cells/100uL and then loading 100uL into each well of a 96-well plate...does anyone recommend any other ways that have worked?

hi
the cells i use and whoch divide themeselves quick (293) shows clones in roughly 2weeks. The method of dilution may be tough. it depends on what number of clones you want to analyze.
Your method seems pretty good but for 30clones you'll need to analyse maybe 3 96well plates.
I switch for an alternative. I pellet 10cm plate cells resuspend in 5ml and pipett 25µl that i put on 10ml on the plate.
Then 2weeks after the plating clones appears.