Hello everyone, two months ago I did a transfection of HEKs cells to insert a miR using the JetPei reagent. On that occasion, the transfection was highly efficient and the cells were in perfect condition. However, now by doing exactly the same the transfected cells get up and die, and the transfection efficiency has dropped dramatically. Also, the control (with JetPei without plasmid) is perfect.
May be you can look at your HEK cells passage numbers. Usually at lower passage number, cells would do better If it was same HEK cells thawed from the same stock.
Did you use a wash to get rid of possible endotoxins into your plasmid prep ?
Enkhtuya Radnaa thanks for your response. The same passage cell were used.
Jean-Pierre Dangy Thanks for your response. I think that my prep kit isn't specific for eliminate endotoxins. But, the plasmid used the second time was exactly the same that the first time, same extraction. And the first transfection worked. The plasmid was frozen between the first and second transfection.
OK great so it's something else. Did you trypsinize the HEK cells ?
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