So would I put the 1.5mL directly into a 15mL conical tube to spin down, or just place the 2mL eppendorf in the larger centrifuge? We have the buckets to do both. I will try extending the time to 20min, and try higher g's. We haven't specifically looked to see what LNCaPs can handle, but that should be straightforward enough, I'll just need lots of tubes!
I use a 200 microliter pipetman to aspirate off about 500 microliters of fluid when I attempt to pellet the cells. I then plate the rest of the volume 200 microliters at a time in the 96-well plate. Results from that, and from just plating the entire volume with no centrifugation (1.5mL), are about the same. Is there a better way to remove supernatant? Thanks for the suggestions, the problem has been really frustrating for me.
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