DNA/RNA random mutation method?

I would like to generate some random mutations on either DNA/RNA,
however, i won't get it with UV mutagenesis method. Does anyone know
any good method to generate random mutations? Thanks a lot!

In article <[Only registered users see links. ].net> , "Z.L. K" <[Only registered users see links. ]> wrote:

Error-prone PCR. Many manufactures sell appropriate polymerases.

DK

On Wed, 13 Jun 2007, DK wrote:


How about hydroxylamine treatment:

[Only registered users see links. ]

Or EMS:

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Use of a mutator strain of _E. coli_ is also an option.


Or, i hear, you can just use the worst normal polymerase you have around
(Taq is fine) and throw in more magnesium, some manganese ions, extra
nucleotides and extra polymerase:

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Or, for Quebecois style EP-PCR, some alcohol, bien sur:

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tom

--
Tom Anderson, MRC Laboratory for Molecular Cell Biology, UCL, London WC1E 6BT
(t) +44 (20) 76797264 (f) +44 (20) 76797805 (e) [Only registered users see links. ]

In article <[Only registered users see links. ].net> , Tom Anderson <[Only registered users see links. ].uk> wrote:

And also drastically reduce concentration of one of the dNTPs.


Yep, that's the ticket. Alcohol is a good way of producing aberrant
progeny :-)

Problem with all of the above is that mutations will not be perfectly
random. Some recent polymerases, OTOH, were tweaked to make
nearly random mistakes.

DK

With the cost of oligos so low I would consider total synthesis with
doped oligos. Probably the only way to control the process well.

Roland

>Problem with all of the above is that mutations will not be perfectly

RNA viruses achieve a highly random mutations ... I wonder if there a
mutagenetic technique involving transcription to RNA, mutation and
reverse-transcription afterwards...?
Erm, before cliking send, I typed it into google and found this:
[Only registered users see links. ] an RNA-based mutation strategy
based on Qb replicase.

I still wonder that, given that mutagens should be more effective on on RNA
than on DNA, why people don't rather mutate RNA...?

max

Oh my dear, they bathed the pol in urea, isopropanol, propan-1-ol, and
butan-1-ol! It must have got quite a hangover! My pol prefers a
delicious Bordeaux Barrique...

For those who want to get access to the paper: The link in PubMed
seems not to work, you'll get an "unknown doi" error message. Check
here directly: [Only registered users see links. ]

Wo