how making site-directed mutation on genome DNA

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seahow
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how making site-directed mutation on genome DNA

Post by seahow » Thu Apr 01, 2010 1:54 am

I want to site-directed mutation on genome other than plasmid DNA. Anyone has protocol like this. I donnot want to introduce neo etc antibiotic maker. I need the clear genetic background which the mutation differ to the wild type with only the nucleotide.
any help is wellcome!!
Thank a lot.

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JackBean
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Post by JackBean » Thu Apr 01, 2010 8:19 am

what about, if you shared the species, you want to mutate? That would be a little simpler to give advice... :roll: :-O
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

seahow
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Post by seahow » Sat Apr 03, 2010 3:00 am

JackBean wrote:what about, if you shared the species, you want to mutate? That would be a little simpler to give advice... :roll: :-O


I donnot understand your advice. I mean just mutate the nucleartide with chromatin status unchanged.
and actually I wanna research the sequence on chromatin in vivo.

please give your advice in detail. thanks!

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MrMistery
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Post by MrMistery » Sat Apr 03, 2010 2:45 pm

well first you have to figure out what your target sequence is (the sequence of the chromatin you want to mutate). Then make an artificial oligo that has the mutation, and introduce it into the genome by recombination insertion.
"As a biologist, I firmly believe that when you're dead, you're dead. Except for what you live behind in history. That's the only afterlife" - J. Craig Venter

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