Protection of DNA During Prep. Agarose...

Angel Yanagihara angel at pbrc.hawaii.edu
Mon Jan 20 21:06:37 EST 1997


Aloha,

I am interested in anyone's experience in replicating the results reported
in:
	Protection of DNA during preparative agarose gel electrophoresis
	against damage induced bny ultraviolet light(1996)
	BioTechniques 21(5):898-903

To summarize, this report indicates a 2 to 3 order of magnitude
enhancement in biological activity (as assessed by transformation
efficiency, PCR amplification or in vitro transcription level) of gel
purified restriction fragment-derived ligation products (using standard
Ethidium Bromide stain-UV protocols) by the inclusion of cytidine or
guanosine in the running buffer at 1-10mM.

If this works, this would be a fantastic advantage for multiple step
ligation protocols.  Is this protective effect depedent on inclusion of
the ribose containing nucleoside or also possible with the cytosine and
guanine bases?

Thank you in advance for any comments on this topic.

Angel
angel at pbrc.hawaii.edu




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