something similar happened before in the lab.

we were using lipofectamine, suddenly the transfection did not work, order a new one, worked.

Hello Peter Wolfrum ,

According to me, you need to tackle this point by point.

First, if you have the possibility to thaw a fresh vial of your cell line, I would highly recommend to try this. In fact, maybe your cells were affected by temperature, humidity or Co2 drop that wasn't under your control. Also, they are maybe contaminated. In my opinion, I think that it is most probably your cells since there's lower chances that both your plasmids or your reagents have a problem.

If it's still doesn't work on your fresh thawed cell line, I would recommend to test the integrity of your plasmids by measuring the 260/280 and 260/230 ratio followed by a PCR.

Last but not least, it can be both of your reagent that had trouble either by contamination or a problem with the storage. If it's the case, you'll need to order new ones and test them.

Hope it helps,

Maxime.

thanks a lot for your help! Appreciate it!

I also could imagine that it was related to the cells and will continue to evaluate this next week

Best regards,

Peter