Any ideas for substitute for chloroform?
hamel at cc.umanitoba.ca
Thu Mar 16 20:47:48 EST 2000
In <rhubner-1B71D9.23311316032000 at news.uia.ac.be> Roland Hubner <rhubner at gins.uia.ac.be> writes:
Thanks for your reply ... you're always on top of things as usual!
I JUST ordered some 1,1,1-trichloro-2-methyl-2-propanol (which is simply
4-trichloro-n-butanol ... Cl-C-CH2-CH2-CH2OH). I suspect this might have
adequate surface tension so that it won't leak ("easily squeeze/slip")
through lid of Eppy tube (it is viscous enough, being a linear 4 carbon
alcohol), AND be hydrophobic enough to denature proteins AND dense enough
to sediment together with phenol after centrifugation. I guess I want it
all ... cake AND icing, eh? :^)
I'll keep everyone posted ... I expect to have thorough enough results in
2-3 weeks (it's back-ordered for 2 weeks at our local Sigma-Aldrich).
Roland, as for your experience with DNA extracted with bromo-chloropropane
and the DNA becoming brownish after being stored in -20*C for 1-2 years, I
suspect the bromo in bromo-chloropropane might be harmful for RNA & DNA in
the long term. Bio-molecules such as proteins, nucleic acids, lipids,
steroids, etc don't cope well in presence of brominated organics.
I wonder what would happen if Piotr Chomczynski substituted the alcohol
form sans bromine (with an extra chlorine instead)?
I'm gonna email Piotr Chomczynski to get his 2 cents worth.
>> Does anyone have any practical experience using a substitute for
>> chloroform during nucleic acid extractions?
> Piotr published this one a while back:
>/Chomczynski P, Mackey K
>/"Substitution of chloroform by bromo-chloropropane in the single-step
>/method of RNA isolation"
>/Anal Biochem 1995 Feb 10;225(1):163-4
> Well, one sample of Mycobacterium smegmatis RNA that I store at -20?C
>turned brown after a couple of month... never took time to inspect...
More information about the Methods