prepare the SCLC for the transfection - SCLC cell seeding for transfection (Apr/23/2008 )
I am used to work with 293T cells but i need to seed SCLC cells overnight to reach 80% confluency in 6well plate for trasfection. I am actually making stable clone.
so far i dopnt knoe hoe much cells to plate and when i plated last night there too many clumps even though i did try to resuspend to like 10mins.
Some cells are hard to get off the plate. I usually wash twice with trp-EDTA, instead once. Increase the concentration of Try-EDAT to 2x is another possibility. There is 10X Trp-EDAT that you can buy.
thanks, but my main problem is to determine how much cells yields 50% or 80%
let your experiment to determine the condition. different cell type has different cell volume and growth rate, therefore, its hard to ask someone to predict it for you. Do a cell counting, seed different amount of cells according X cells/ml in 2 ml volume, and find out by yourself. You will get answers in 24 hrs.