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How to make tris-saturated chloroform

DNA purification plasmid chloroform

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3 replies to this topic

#1 k_undertoe



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Posted 30 October 2012 - 08:41 AM

So, I've seen several recommendations when doing phenol:chloroform extraction to first use a saturated phenol:chloroform:isoamyl alcohol solution followed by a second extraction using only chloroform. I have the correct buffered and saturated PCI solution, but now I am at a loss: what exactly constitutes a 'tris-saturated chloroform' and more importantly how do I make it? I have now some pure chloroform (well, alcohol added as a stabilizer...) but what do I need to do to it in order to use it in my extraction? The specific product I have is this: spectrophotometric grade ACS reagent chloroform, Sigma 366919 (C-5312)

Edited by k_undertoe, 30 October 2012 - 08:44 AM.

#2 hobglobin


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Posted 30 October 2012 - 08:48 AM

Actually I never did it but the recommendations from protocols (and many bioforum posts) all have the same content: buy it as ready-made mix as this doing it yourself is a bothersome and not that easy process. And the finished stuff is cheaper too.

One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.

#3 phage434



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Posted 30 October 2012 - 10:35 AM

Chloroform has no acid/base groups, and is largely insoluble in water. You can use chloroform without further treatment or pH control, unlike phenol-chloroform or phenol.

#4 BMF



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Posted 03 November 2012 - 08:24 PM

I have never heared that until now.
Maybe you mean saturating phenol with Tris (pH 7.9 for DNA and pH 5 for RNA)


Edited by memari, 03 November 2012 - 08:25 PM.

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