[DIYbio] Re: DIY targeted evolution

T->U will cause A to replace the U in transcription.
http://www.sciencedirect.com/science/article/pii/S1097276503003605

Things like alanine GCT->GCA causes no change and glycine GGT->GGA and many others will cause no change, 27 codons don't even have a T. You will mutate your start sites (ATG) and your top codons(T**). Your search space is so small using this technique because each codon will only have at most(TTT) 3 other possible codons it can become. So you will be able to try 1000s(10^3-10^5) of different constructs. Not a large search space. Most mutagenesis strategies search 10^9 - 10^12 sequence space and are limited by transformation efficiency.

Random mutagenesis has been done so much just look up papers.
A simple way is to make degenerate primers.
Or use a MutL or MutS deletion strain of bacteria.
or directed evolution
http://nar.oxfordjournals.org/content/28/16/e78.short

or many many other things.

http://scholar.google.com/scholar?hl=en&as_sdt=0,14&q=bacteria+random+mutagenesis

On Monday, August 19, 2013 2:15:20 AM UTC-5, Mega [Andreas Sturm] wrote:

Interesting...


But I guess it's gonna be like that:

I ask a chemist friend to produce HNO2 in situ. Then I incubate the plasmid in it and do a transformation.


Maybe that'll be sufficient to produce a lot of mutants....