For a Luciferase, I was harvesting some Hek293T cells.
I had added 1 ml of PBS per well, and then collected the wash, centrifuged it down, and pipetted the supernatant out. I then added 200ul 1x RLB to the cells in the wells. I was then called away to learn how to thaw out new cells, so the Hek293T's I had just added RLB to ended up sitting for 45 minutes. When I came back, they were a strange viscous goo consistency. I transferred the goo to their respective PBS counterpart, and let it sit for 20 minutes. The cells were then spun down, and the clarified liquid taken off and put into fresh tubes. I used this to run the assay. My numbers were very very low.
I was trying a new method of harvesting... how many ways did I go wrong? (Please be gentle...)
Reporter Lysis buffer exposure time
No replies to this topic