sources of agarose affect transformation

Keith Rand Keith.Rand at molsci.csiro.au
Tue Aug 11 19:25:21 EST 1998


In article <6qnq6m$8k9$1 at nntp.ucs.ubc.ca>, lpss at unixg.ubc.ca (Alex Chang) wrote:

> I used to get very good results using BioRad agarose for
> electrophoresis, Qiagen gel extraction, ligation and transformation. 
> 
> The lab I am in now are using GIBCO/BRL ULTRA PURE agarose for 
> electrophoresis. My transformation assays have been less efficient 
> since I came to this lab.
> 
> I am wondering whether others have similar experience with GIBCO's 
> product?
> 
> Alex

I havn't had a problem with that agarose.

Presumably you are now using a different transilluminator. If so, could
you try exposing a small amount of whatever plasmid dna that you are using
as vector to the uv whilst you cut out bands? If you find that the dna is
damaged by the uv (as is generally found when people do this test) then
you could consider avoiding uv exposure altogether. Of course, the amount
of damage depends on many factors, but especially the amount of the
shorter wavelengths of uv that come through the filter. I have a method
that uses crystal violet that stains the dna bands during electrophoresis
if you think this might help.

-- 
Keith Rand,  Sydney Australia



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