coady at ERE.UMontreal.CA
Wed Nov 9 14:23:20 EST 1994
In article <jpcd0-0811941439180001 at macr1-2.welc.cam.ac.uk> jpcd0 at mole.bio.cam.ac.uk (John Dixon) writes:
[Much deleted- I've replied via email, on some topics that have
been considered here before, except for........]
> I am planning to use 900ul of lysate, 90 NaAc (2M ph4.0!), 900 Phenol
>(ph4.2 DEPC H20 equilibrated) and 100ul Chloroform:Iso-amyl alcohol (49:1)
>which should just squeeze into a 2.2ml eppendorf so that I can spin my
>samples at the reqd 10000g in microcentrifuges.
Something that I learned the hard way is that you never do
phenol/CHCl3 extractions in 2.2 ml tubes; we usually found that
they would develop a small hole at the base and all of the contents
would end up decorating the inside of the centrifuge. This might
vary between different manufacturers, but I doubt it; I'd phoned
one of them at the time to inquire about this and he thought that
it had to do with how they were bonding the bases of these tubes
to the sides.
I've seen a number of people get angry when they do this
without knowing that the tubes will leak, so avoid this.
Michael J. Coady
COADY at ERE.UMONTREAL.CA
The opinions expressed above are solely those of the author and do not,
in any way, shape or form, represent the Universite de Montreal.
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