I am trying to find a pair of potential enzymes that would create compatible sticky ends for a protein fusion. I am very limited for enzymes that I can use. is there any site or program that I can use that would allow me to enter the name of an enzyme and give me the one that would make a compatible sticky end to it. I tried NEBs but it kept giving me wrong results.
Hi there,
You get this info either from NEB catalog (compatible cohesive ends section) or from NEB website:
https://www.neb.com/tools-and-resources/selection-charts/compatible-cohesive-ends-and-generation-of-new-restriction-sites
Hope it helps.
I would strongly recommend you forgo using restriction enzymes entirely and instead use a form of Gibson Assembly (https://en.wikipedia.org/wiki/Gibson_assembly). For example, NEB sell a proprietary kit for performing this kind of assembly: https://www.neb.com/products/e2621-nebuilder-hifi-dna-assembly-master-mix . This only requires you to digest your vector, inserts are not digested and just assembled with digested vector in a one pot reaction. NEB even have a tool to design the PCR primers http://nebuilder.neb.com/ . Say goodbye to restriction-ligation cloning!
Hi Andrew,
Compatible Cohesive Ends and Generation of New Restriction Sites from NEB BioLab would give you the perfect answer.
Website is attached below for your convenience:
https://www.neb.com/tools-and-resources/selection-charts/compatible-cohesive-ends-and-generation-of-new-restriction-sites
Best,
Hao
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