I am trying to optimize Nucleofection to MEFs using home-made buffers, since the buffers from the company are expensive. Could anyone please help me with this? I would be very grateful if you did.
One option is to use either RPMI1640 or DMEM media without serum and antibiotics and use A20 or L-17 program It will give you reasonable nucleofection. Also the cuvette can be washed with ethanol and reused. This was tested by one postdoc in our lab with GFP expressing plasmid. You need about 2mcg and 3.0E6 cells at least.
Just in case nucleofection efficiency is an issue when using DMEM or RPMI the alternate is to use transfection reagents from Mirus Bio. called Ingenio® Electroporation Products. It is much cheaper than Lonza and has same efficiecy. They also offer free sample to try.
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