We got almost 100% transfection efficiency by Lipofectamine 2000. Now it is low, probably only 30 to 50%. Just wondering if the cells have been contaminated with mycoplasma, affecting the transfection efficiency. Any suggestions and experience? Thanks.
probably u r rit...either the cells are contaminated or stressed. I faced similar situations with several cell lines where the transfection efficiency got reduced due to contamination.
probably u r rit...either the cells are contaminated or stressed. I faced similar situations with several cell lines where the transfection efficiency got reduced due to contamination.
Hi Jia-Wang,
In my experience, simply a high passage number of the cultured cell lines can negatively affect transfection efficiency as well as cell death rates during transfection etc. Try taking a new batch of low passage HeLa cells to culture and test whether this helps. More useful tips can be found for example here:
http://www.lifetechnologies.com/fi/en/home/life-science/cell-culture/transfection/transfection-support/transfection-tips-and-tricks.html
That's quite a drastic change in efficiency, so you're probably on the right track thinking it's contamination. If there's a problem with your culture, you may just want to order a new batch of cells. Otherwise it might be the transfection reagent having been altered in some way. HeLa isn't that difficult to transfect (we use this kit: https://altogen.com/product/hela-transfection-reagent-cervix-adenocarcinoma-cells/) and I've never really had the same kind of changes that you've had.
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