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For interfacial studies, I would like to build an alanine scanning library for one of my proteins examining 20 sites. I will ultimately express the gene using E.coli cell-free protein synthesis. I already have the template gene which was originally built using PCR assembly. Now, what's the best way to build a scanning library?

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do you mean, how do i decide which amino acids to include in the alanine scan or how do i create the library of alanine mutants? – shigeta May 17 '12 at 19:54
@shigeta, definitely more "how". – bobthejoe May 17 '12 at 21:28
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genscript.com/alanine_scanning.html genscript has some pages to help you reason through the design. Myself I'm not sure, but it looks like it takes quite a lot of oligos to say the least... – shigeta May 18 '12 at 6:08

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