My experimental steps are as follows: transformation, isolation, transfection, titration.
In the titration I set up in the last 3 experiences, my jurkat cells do not gather in the middle. When checking jurkat cells with flow cytometry after titration, my 10ul and 3ul cells are very unhealthy, while the results of other concentrations are very low. I also prepare control GFP alongside my variants. In them, my jurkat cells are scattered, but my flow results are normal. I also did it using pspax and vsvg isolated at different times and different jurkat cultures to understand why, but the result is still the same. I use hek cells for transfection and see GFP glow in the fluorescent microscope. I think that if my transfection had failed, I would not have seen the glow in the fluorescence microscope and the control gfp would not have come out normal. Is it true? What could be the reason why the cells look like this, are unhealthy and my flow results are low?
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