Random Mutagenesis

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lcronan
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Random Mutagenesis

Post by lcronan » Tue Jan 20, 2009 10:11 pm

I am attempting to make a library of copy-number mutants. In order to do this, I wanted to use random mutagenesis to create the mutations. Does anyone know of a good, tried and true technique? I have tried a random mutagenesis kit, but so far have gotten no mutants. Of course, I may be doing something incorrectly, but I am not getting any helpful feedback from my boss... and yes, I have asked! If anyone has any advice, hints, knowledge, thoughts, etc. to share on the subject, I would be extremely grateful!

Thank you!
Leah

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mith
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Post by mith » Tue Jan 20, 2009 10:44 pm

what kit?
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Enjoying one moment at a time;
Accepting hardships as the pathway to peace;
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lcronan
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Re: Random Mutagenesis

Post by lcronan » Wed Jan 21, 2009 11:38 pm

Stratagene's Random Mut. Kit

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Re: Random Mutagenesis

Post by lcronan » Mon Feb 02, 2009 6:54 pm

OK, I found a method that works:

Hydroxylamine Mutagenesis. I got sooo many mutants, now it's time for screening...

I tried one protocol that asked for incubation at a high temp for a short amount of time. This didn't make sense to me since hydroxylamine is denatured at high temps, but I tried it anyway and got no results. I tried another that asked for incubation at 37 for 20hrs and that seems to have worked beautifully!

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