Boiling Method for Minipreps

jchen at CALSHP.CALS.WISC.EDU jchen at CALSHP.CALS.WISC.EDU
Sat Jul 31 00:55:11 EST 1993


> 
> I would like to know if anyone uses the boiling method for plasmid
> minipreps to prepare DNA for sequencing.  I wish to try this method for
> several pBR322 constructs I have recently transformed into HB101.  Any
> hints would be appreciated.
> ^X
> 
> 
I have been using boiling method to miniprep DNA for sequencing and it has
been successful.  Prepare plasmid (I used pUC) with standard method. 
Phenol extraction is not necessary.  Denature DNA with NaOH following
the procedure  of using Sequenase (2.0).  In the case of not getting enough
DNA, simply use 3.0 ml (spin twice), rather than 1.5 ml of culture.  pBR322
will probably work the same way even though I didn't try it.

Good luck

J. Chen
jchen at calshp.cals.wisc.edu
Department of Food Microbiology and Toxicology
University of Wisconsin




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