defrosting........how do you do it? - (Aug/07/2007 )
When defrosting molecular biology reagents (such as primers) is it best to either;
A) Defrost them on ice. Or,
Defrost at room temperature and then work with them on ice after they have defrosted…?
Any opinions will appreciated!!!
I have not noticed any difference between the two. And I would guess that there is little long term difference between the two methods.
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.
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.
I also did like what has been mentioned by scolix
I was taught to do them in water at room temp using floats. I have been doing it in my hand like Scolix.
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.
armpit DNA (aDNA?)
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
- writing this post has just turned me from enthusiast to veteran so i must be right
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.
I stick the tubes into my glove, so they're nice and worm. I only put them in ice after thawing.