Consistent subcloning from agarose (finally !)

Hugo Eric eric at bcserv.wustl.edu
Tue Apr 6 05:45:39 EST 1993


pnh at fcsparc6.ncifcrf.gov (Paul N Hengen) writes:

>jgraham at bronze.ucs.indiana.edu (the End) writes:

As part of a followup to these two methods of subcloning agarose
gel purified DNA fragments I have found that using methylene blue
to visualize the bands vs. EB UV light results in a 10-fold higher
efficiency in insert-containing clone isolation.  Simply stain
your agarose gel in 0.25% methylene blue in 0.1X TAE for 30 min
and destain in 0.1X TAE until you can see the band you want.
The only contraindication I know for this method is that it doesn't
work with a product called Synthagel.
                                   Eric
-- 
  Eric R. Hugo, Postdoctoral Research Associate |eric at bcserv.wustl.edu
  Dept. of Biochemistry and Molecular Biophysics|                
  Washington University School of Medicine      |               
  Box 8231, St. Louis, MO 63110_________________| (314) 362-3342



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