basics

Basavareju Shankarappa bsh at MED.PITT.EDU
Wed Jul 15 11:32:45 EST 1992


>re:
>I'm trying to set up a "dsDNA cycle sequencing"
> protocol...

Paulo

>Heck! What are the basics? Does the average molecular biologist
> know why you use 2 vols of ethanol to ppt DNA, but only 1 vol of
> isopropanol? or why monovalent cations aid the pptn? Or the chemistry
>Mike "I'll use any kit, any time" Poidinger
>Dept of Microbiology
  >University of Reading


Well, I am interested in knowing the difference between two vol of
ethanol and one vol of isopropanol.

I have observed that using NaCl for pptn of DNA often results in 
precipitation of salt specially after prolonged incubation at 
-20 C.  I learnt this from my experience rather than anything.
I have also learnt that this salt pptn is very very hard to get rid off.
Does somebody know answers for the above questions?

Will it be a good idea if people periodically post some of the 
simple yet unique resons behind the crazy procedures we use. 
I think all of us might learn something

Do I hear anyone second my opinion?

Raj  Shankarappa




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