Chomczynski traumas!

Simon Dawson mbxspd at unicorn.nott.ac.uk
Wed Nov 9 07:36:18 EST 1994


Ok....an answer to the 2M NaOAc pH4...maybe. We have had the same
problem. Pain in the *** isn't it? The way we do it now is...say we
need 100ml, once all the NaOAc solid is weighed out, I added a small
amount of H2O...not minimal to dissolve it, just a small amount. I'm
not really sure how much...you might have to experiment :)
Then, we basically dissolved the solid in glacial acetic acid. It goes
a bit gooey when you do this as I recall, but it was the only way we
didn't end up with more volume than we were supposed to have!
I suppose basically you could just add glacial acetic acid to the
NaOAc solid straight away but I imagine this might be really gooey.
If it did dissolve like that, you can then just make the volume up
at the end with H2O.
You might also try warming the solution a bit.
I can't recall exactly how we made it up last time, but it was
something like that :P
As to the other questions, we still use the '86 method. As it worked
so well we never saw the need to change it :)
We dont ever scale down volumes as mentioned. Someone here did and
they found that they ended up with DNA contamination in their
final RNA sample.

Simon Dawson
Dept Biochemistry
Queens Medical Centre
Nottingham
UK

Internet email: mbxspd at unicorn.nott.ac.uk



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