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Does autoclaving destroy DNA

Does autoclaving destroy DNA - Protocols and Methods Forum

Does autoclaving destroy DNA - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 02-21-2007, 11:37 AM
newsnet customer
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Default Does autoclaving destroy DNA



Hi,

Does autoclaving (121C for 20mins) DNA destroy it? What I mean by "destroy" is to break the covalent bonds between adjacent nucleotides and NOT the breaking of ionic bonds between the two strands. I argue that 121C is well above the melting temperature of DNA (95C), which should do the trick.

Keep your answers short and to the point please.

Cheers,
ST
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  #2  
Old 02-21-2007, 02:54 PM
jg374@cam.ac.uk
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Default Does autoclaving destroy DNA

>

No. Although it break a lot of it into fragments <100mers. The melting
temp refers ot the inter-strand bonds so is not relevant.

[Only registered users see links. ]


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  #3  
Old 02-21-2007, 09:56 PM
WS
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Default Does autoclaving destroy DNA

Suggestion for an experiment:

autoclave an eppi with say 10ug of your favorite plasmid and then
transform the solution into your favorite bacteria or PCR for the
insert.

Wo

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  #4  
Old 02-21-2007, 11:27 PM
Jose de las Heras
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Default Does autoclaving destroy DNA

Years ago, we used to autoclave wheat DNA as a substitute for sonication...
It doesn't work as well as sonication, but you do get a smear in the end.
I never tried it on short molecules, like plasmids etc, so I don't know how
well those would fare.

Jose


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  #5  
Old 02-22-2007, 12:52 AM
DK
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Default Does autoclaving destroy DNA

In article <1172094972.016487.106760@t69g2000cwt.googlegroups .com>, "WS" <[Only registered users see links. ]> wrote:

Prediction: if the solution was non-buffered, expect 100X drop
in efficiency. If the pH was >7.5 at RT, expect only a 10X drop.

DK
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