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That is DNA breathing? - (Dec/05/2011 )

That is DNA breathing?

-yqluuf-

Its the "opening" of the DNA.., forming small "bubbles" .. its when the nucleotides are a bit away" from each other.
You should find some information about it using google.
Just type in DNA breathing...

-pito-

Thank you pito, After fill in your DNA fragment by klenow, you heat inactivate klenow. According to the NEB website, you should add EDTA to protect the DNA ends as they "breathe" while the temperature is increased. My question is, after inactivating, the opened DNA should anneal together, if so, I can’t see any problem can be caused by DNA “breathe”.

-yqluuf-

yqluuf on Tue Dec 6 18:54:21 2011 said:


Thank you pito, After fill in your DNA fragment by klenow, you heat inactivate klenow. According to the NEB website, you should add EDTA to protect the DNA ends as they "breathe" while the temperature is increased. My question is, after inactivating, the opened DNA should anneal together, if so, I can’t see any problem can be caused by DNA “breathe”.


I think what they mean is that you need EDTA while you are heating the samples. During the heating the DNAses are not inactive right away.
And because the DNA breaths during the heating, its more "open" to the DNAses..

when the DNASes are inactive after heating, then yeah, it should not matter of they breath.

(well thats what I think, but I am not really an expert on DNA, maybe its best to open a new topic with this specific question in it)

-pito-

Thank you.

-yqluuf-