I am facing problem in setting up in vitro translation reaction. It looks like I need an energy generating system like phosphoenol pyruvate and pyruvate kinase . There are kits available (NEB) also, but they are pretty costly. If I prepare cell extract and then use it for IVT will there be any problem? How much does omitting energy generating system affects the reaction? I have came across protocols for IVT but all uses the PEP and pyruvate kinase, is there any way to bypass it by using excess ATP?

Thanks in advance.

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