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Effect of freezing on DNA methylation?


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6 replies to this topic

#1 egerecske

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Posted 28 January 2009 - 11:24 PM

hi everybody!

Well, it was a bit surprising to me that my account doesn't exist but ok, now I understand why.
So my question is: do you think it has any effect if I freeze cells before izolate DNA for BS studies...? Can freezing change methylation status of promoter? I haven't read anything about it but maybe it is important...

Cheers

#2 pcrman

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Posted 03 February 2009 - 09:04 PM

I don't think freezing cells or DNA changes its methylation, but it is not absolutely impossibe.

#3 rkay447

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Posted 03 February 2009 - 09:58 PM

I don't think freezing cells or dna significantly alters the methylation state. I've tried to digest previously frozen vector with an enzyme inhibited by methylation and the digest did not work. (oops)

#4 toejam

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Posted 10 February 2009 - 02:21 AM

it has been reported that there's an increase in DNA methylation in plants that undergo dormancy. if you're working with plants might find this review useful:

Knowing when to grow: signals regulating bud dormancy
David P. HorvathE-mail The Corresponding Author, James V. Anderson, Wun S. Chao and Michael E. Foley
Trends in Plant Science
Volume 8, Issue 11, November 2003, Pages 534-540
"When there's no more room in hell the dead will walk the Earth"

#5 et2b

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Posted 02 March 2009 - 06:33 AM

it has been reported that there's an increase in DNA methylation in plants that undergo dormancy. if you're working with plants might find this review useful:

Knowing when to grow: signals regulating bud dormancy
David P. HorvathE-mail The Corresponding Author, James V. Anderson, Wun S. Chao and Michael E. Foley
Trends in Plant Science
Volume 8, Issue 11, November 2003, Pages 534-540



hmm saw this same question somewhere else on the forum....but again :rolleyes:

if your cells are death and the protein is denatured, thus inactive......your cytosine methylation is OK. Its a covalent modification of DNA, quite stable.

#6 Trof

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Posted 09 March 2009 - 03:07 AM

We have a similar problem here, we want to analyse few years old -20 stored DNA for methylation pattern. Is it still usable?

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

'Normal' is a dryer setting. - Elizabeth Moon


#7 toejam

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Posted 09 March 2009 - 03:28 AM

Is it still usable?

i would say there is only one way to know it....
if the samples haven't been thawed it should be fine.
"When there's no more room in hell the dead will walk the Earth"




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