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defrosting........how do you do it? - (Aug/07/2007 )

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When defrosting molecular biology reagents (such as primers) is it best to either;
A) Defrost them on ice. Or,
cool.gif Defrost at room temperature and then work with them on ice after they have defrosted…?
Any opinions will appreciated!!!
Thanks,
Eve

-evejoss-

I have not noticed any difference between the two. And I would guess that there is little long term difference between the two methods.

-perneseblue-

i think that thaw on ice is the best method recommended.
But a pernesblue, i didn't see ig difference.

For proteins(extracts or purified ones), RNA, NTP, i thaw them on ice of course.

-fred_33-

I usually thaw them in my hands and keep the primers on ice once completely thawed. As long as they are not contaminated, they seem to work fine.

-scolix-

I also did like what has been mentioned by scolix

-money-

I was taught to do them in water at room temp using floats. I have been doing it in my hand like Scolix.

-Bungalow Boy-

For robust things such as PCR buffers, MgCl2, etc., I put the tubes in my armpits. Nice and warm (faster thawing) and it leaves my hands free to do other things. I don't work with human material, so the worry of contamination with armpit DNA hasn't been an issue.

-wbla3335-

armpit DNA (aDNA?) huh.gif
because these things (aliquoted antibodies too) are in such small measures they usually take care of themselves after a minute - i doubt that either way will make much of a difference

dom

- writing this post has just turned me from enthusiast to veteran so i must be right

-Dominic-

I'm a fan of laying the tube on ice so that one side is touching the ice, the other is at room temp. If I have time I thaw on ice like a good scientist, but most of the time I just let it thaw at room temp then put it on ice as soon as I can. DNA is rather stable if stored in TE or miniprep elution buffers. Primers are stable in TE as well, so I usually store those at 4degC, so no thaw necessary. Same for any DNA plasmids and fragments I know I will be using alot in the near future.

-smitity-

I stick the tubes into my glove, so they're nice and worm. I only put them in ice after thawing.

-swanny-

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