Simultaneous saturation mutagenesis using FlexDDG protocol

[yashrajk]

Hi,

I have used the protocol to do saturation mutagenesis at each residue position. However, I am interested in performing simultaneous (double or triple) mutations using the protocol and then obtaining the most preferred set of two/three residues.

I feel this is important because the adjacent residues will affect the stability of a sidechain. Currently, I am performing point mutations on a poly-GLY peptide, and am getting Trp or Phe as the best AAs for each position. It will be impractical to have 3 or 4 consecutive Phe or Trp residues.

Can you or anyone among the users help me out with this? Maybe it's a simple edit to the script or generating a new .resfile?

Thanks,
Yash