Philipp,
You say essential residues, which I understand that you have an idea of the
protein's function. If so, I would look at the known functional motifs and
mutate the residues in those first. You can also try using those protein
structure prediction tools from expasy and identify structurally important
regions (strands, helices, etc.), instead of just mutating every residue.
You might also be able to identify the residues that form contacts with each
other, meaning that those contacts might be important for maintaining
structural integrity of the protein. Terminal residues would probably be
less important that internal ones. Etc., etc. This is just a suggestion.
Hope it is helpful, although it is not exactly the answer to your question.
Cheers,
Emir
"Philipp Wechner" <philipp.wechner at uibk.ac.at> wrote in message
news:3D173F3A.F687E222 at uibk.ac.at...
> Hello to all of you.
>> I have a protein-coding sequence of 600 bp and I would like to random
> mutagenesis with this sequence to find essential residues of the
> Protein.
>> does anybody have experience with random mutagenesis kits? I know the
> Diversify PCR Random Mutagenesis Kit from clontech, but I never tried
> it. Do you know how it works and how many different mutants you can get
> with it? Are there better methods and kits around?
>> Any help is welcome.
>> greetings
>>
