any ideas for substitute for chloroform

"Hamel, Andre AGR AHamel at agr.gov.mb.ca
Fri Mar 24 11:40:35 EST 2000


Greetings bionetters,

	Last week I rec'd the trichlorobutanol (Sigma-Aldrich cat no
11,205-4) and
tested it on several preps (DNA and RNA from tissues, blood, semen and
serum).
They all looked normal and PCRed great (no changes from nucl acid using
CHCl3).

	My preliminary findings are that it works GREAT as an additive in
chloroform!
The viscosity and surface tension are dramatically increased, yet all the
denaturing and phase
separating properties of chloroform are retained.

I wonder how come nobody ever tested this before (maybe not commonly found
on shelf)?

1,1,1-trichloro-2-methyl-2-propanol (trichlorobutanol, herein referred to as
"TCB") 
has a melting point of 77*C. 

I dissolved 25 grams of it in 100 mL of chloroform (at room temp).

After shaking or stirring, the TCB dissolved nicely in the CHCl3. It stores
okay in fridge (1 week so far
and no visible ppt) ... in -20*C it turns cloudy, but brief shaking turns it
clear again.

For nucl acid ext'ns I simply substituted this TCB(25% w/v)-CHCl3 for
CHCl3-2% isoamyl in my extractions 
(GiTC, phenol/CHCl3, etc).

No more CHCl3 flying out of pipettors due to its lack of any surface
tension.

No more leaking out of Brinkmann-Eppendorf 1.5 mL polypropylene microtubes
during vortexing.

The TCB is borsch cheep ($50 Cdn for 500 gm ... adds only a penny per prep)

cheers,

Andre Hamel
Scientist
Virology Laboratory, Veterinary Services
Manitoba Agriculture
Winnipeg, Manitoba  CANADA

email: ahamel at agr.gov.mb.ca

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