Hello,
Is it necessary to change culture medium before transfection. My cells abruptly lose attachment upon medium change.
Thanks.
When do you plate the cells? Mine are plated 2 days before transfection and I do not change media. If you plate them further in advance, then you might have to change media. I do add fresh media the day after transfection to keep the cells happy.
We don't change media as well. But do take note that antibiotics might be bad for cells during transfection. We do not use antibiotics in culture media during transfection process.
It's depend on the transfection reagent you use. In some cases you can just add the transfection mix to the regular growth media, in other case you need to have the cells in media without antibiotic or without serum.
Hi, Just take a look at a few points I have mentioned
1. What cells (Normal Vs Tumor) are you working with? Because I have observed that normal cells are not attached to the substratum since they are not efficient in generating the ECM components themselves compared to those of tumor cells. You could always pre-coat the flasks or plates with 0.2% porcine skin gelatin in H2O.
2. Additionally what confluency the cells are at the time of transfection? Typically it should be around 70% confluent.
3. Like Yi Jet Tan mentioned it's not a good idea to add antibiotics during transfection related experiments.
4.Assuming that your problem is pre-transfection, i would suggest you to change the medium in which the cells are being grown or play around with the concentration of some of the contents.
5. Additionally, some sort of cryptic contamination (like that of mycoplasma) also lead to the type of consequences you have mentioned. Check for contamination as well.
Hope it helps!!
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