The mutation can be inserted via the corresponding side-directed mutaion assay kit. I would propose to check the manufacturers protocoll for such a kit.

Hi @Andreas Eisenreich

Thanks to reply!

I didn't know I could use a kit to perform that. I designed all my primers (WT and mutated), is there any way to perform the assay without this kit? 'Cause I've checked some papers and not all of them say that they used a Kit for it.

You can use kit or other techniques.