How to increase the frequency of mutation by using GeneMorph II Random Mutagenesis Kits (Error prone PCR/directed evolution)?

23 May 2016 1 8K Report

Is there any conventional methods to introduce mutation in lipase gene to make it thermostable? Any effective protocols for gene cloning and transformation after Error prone PCR?

Arunangshu Das

The best option so far I know is to perform the er-pcr and ligate it to your vector of choice followed by electroporation. Electroporation is vital because of its high transformation efficiency which makes library size large increasing the probability to get the best performer. How ever the trickish part is to develop a proper screen for the mutants that is easy fast reliable to find out the best mutant.

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