I already have my restriction enzymes from my gene of interest. The question is, how do I get the correct plasmid for this? I am struggling with all the donor plasmids I see on the internet because they do not have the same restriction enzyme in their multiple cloning sites. Any tips?
If you posted what restriction sites your GOI has, then maybe someone could help you find a plasmid.
But the easiest thing to do would be to design PCR primers to amplify your GOI and introduce useful cloning sites for your vector at the ends of the PCR primers.
1. Use primers (PCR) to create restriction enzyme site on each end of your Insert to fit the vector
2. Use seamless cloning techniques ,ex: Gibson cloning
3. Use Ligation independent cloning (LIC), ex SLIC
https://bitesizebio.com/22117/say-goodbye-to-restriction-enzymes-and-ligases-an-introduction-to-sequence-and-ligase-independent-cloning-slic/
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