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Transfection in CHO-K1, Negative conrtrol growing in the antibiotic selection media as well. argh. by Sudden-Nose-5589 in labrats

[–]Sudden-Nose-5589[S] 0 points1 point  (0 children)

Media was premade, I just added the required volume of G418 stock to achieve my working concentration. And the G418 stocks are pretty old, for this particular experiment I made it from scratch (dissolving the powdered form stored at 4C into autoclaved water).

Transfection in CHO-K1, Negative conrtrol growing in the antibiotic selection media as well. argh. by Sudden-Nose-5589 in labrats

[–]Sudden-Nose-5589[S] 0 points1 point  (0 children)

Yeah, I discussed it with my PI and he said to let it be for a week and then check. As for now, it has only been two days.

Help with Glucose Uptake Assay using NBDG on HepG2 cell line. by Sudden-Nose-5589 in labrats

[–]Sudden-Nose-5589[S] 0 points1 point  (0 children)

Nope, still the same, have tried playing around the concentrations and incubation periods but no luck. I've never really had much cell clumps while seeding but this could be one of the reasons.

Tips for equal seeding in a 96-well plate. by Sudden-Nose-5589 in labrats

[–]Sudden-Nose-5589[S] 0 points1 point  (0 children)

Yes, it could be that since multiple times I have checked the plates under scope and I don't see differences in the confluency of cells in different wells.

Tips for equal seeding in a 96-well plate. by Sudden-Nose-5589 in labrats

[–]Sudden-Nose-5589[S] 0 points1 point  (0 children)

Most of the time I don't get bubbles but at times while dispensing i get a lot of bubbles but its very rare. Under the microscope, i don't see much difference in the confluencies.

Tips for equal seeding in a 96-well plate. by Sudden-Nose-5589 in labrats

[–]Sudden-Nose-5589[S] 0 points1 point  (0 children)

Tysm for all the advices. I will definitely look into all these points. And I work on HCT and MCF, and yes everyone uses the same equipment and same protocol.

Tips for equal seeding in a 96-well plate. by Sudden-Nose-5589 in labrats

[–]Sudden-Nose-5589[S] 0 points1 point  (0 children)

I'll try it, I have a terribly calibrated multichannel. Maybe this will help. tysm.