DNA concentration

Hiranya Roychowdhury hroychow at NMSU.EDU
Mon Aug 21 10:14:55 EST 1995


On 21 Aug 1995 leverone at chuma.cas.usf.edu wrote:

> Giorgio:  yes;  add an equal amount (vol) of butanol to your DNA soln.; 
> vortex; centrifuge; take off top layer (butanol layer).  Repeat this 
> until your DNA soln. has decreased to the desired volume.  Problems with 
> this protocol are:  shearing of DNA during vortexing and remaining 
> butanol in DNA soln.  The latter can be removed by either ppt. the DNA or 
> drawing off some of the DNA soln. when the butanol is aspirated. 
> 
> On 17 Aug 1995, Giorgio Spagnol wrote:
> 
> > Dear fellow scientists,
> > do you know any good protocol to concentrate DNA, apart from
> > precipitating it?
> > Any suggestion would be greatly welcome.
> > Thanks in advance.
> > Giorgio
> > 
> > 
> 


This is a reliable method and also removes any EtBr that one may have in
DNA from a gel. Vortexing is not required. I used to do inversions by
hand. A simple lab rotator (or a platform shaker) will also work fine.
Since the butanol is not water-saturated, it does not take too long for it
to absorb the water..  The residual butanol may be evaporated in a
Speed-Vac or by placing the tube in a dessicator with suction.
			>>>>>>>>>>>>>>>>>>>>>>>>>>>
			  Hiranya S. Roychowdhury
   			  Plant Genetic Engineering Lab.
			  Box 3GL, NM State Univ.
			  Las Cruces, NM 88003
			  Phone: (505) 646-5785
			  hroychow at nmsu.edu
			<<<<<<<<<<<<<<<<<<<<<<<<<<<




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