Having not so much experience with cell-free expression systems, I do have a general question. I am using a commercial RTS 100 E.coli kit to express eGFP. When following the expression on a tecan reader, I observe linear increase of fluorescence instead of reaching a plateau, even with the control vector. Can one tell me whats going wrong?
Hello, Green fluorescent protein is a quantitative reporter of gene expression. The linear increasing of fluorescence means that expression ongoing but finally you reach plateau when expression stopes or GPF expired. For more info see attached info.
Good luck!
Hello, Green fluorescent protein is a quantitative reporter of gene expression. The linear increasing of fluorescence means that expression ongoing but finally you reach plateau when expression stopes or GPF expired. For more info see attached info.
Good luck!
gfp halflife is several hours.. if you run you expression for a few hours you will see only accumulation as new is produced all the time ... so no plateau...nothing is wrong ....... longer time or less ATP or AA in the reaction mixture will help to stop the reaction .. you can add EDTA or stop buffer to see the plateau
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