HELP! -- Protecting DNA on a transilluminator?

Paul N Hengen pnh at ncifcrf.gov
Fri Jul 18 14:23:11 EST 1997


Alex Kukushkin wrote:

> I am having problems ligating my DNA fragments cut out of a gel. I
> get variable results that seem to relate to how long it takes to cut
> out the band. The fastest I can see and cut out a band is about 15
> seconds and by then I suspect there is already significant UV damage.
> Is there any way to protect my DNA during UV exposure?

| In last year there was a report in BioTechniques (21, 898) about DNA
| protection in the agarose gel against UV damage by the addition of
| cytidine or guanosine to the electrophoresis buffer.

There was a very good discussion of that technique some time ago.
The references for it are:

@article{Hengen1997Maytibs,
author = "P. N. Hengen",
title = "Methods and reagents - Protecting vector {DNA} from {UV} light",
journal = "Trends in Biochemical Sciences",
volume = "22",
number = "5",
pages = "182-183",
month = "may",
year = "1997"}

@article{Grundemann.Schomig1996,
author = "D. Gr\"{u}ndemann
     and E. Sch\"{o}mig",
title = "Protection of {DNA} during preparative agarose
gel electrophoresis against damage induced by ultraviolet light",
journal = "BioTechniques",
volume = "21",
number = "5",
pages = "898-903",
month = "nov",
comment = "use 1-10 mM guanosine or cytidine in the gel buffer;
dirk.gruendemann at urz.uni-heidelberg.de",
year = "1996"}

--
*******************************************************************************
* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
* - - Methods FAQ list -> http://www-lmmb.ncifcrf.gov/~pnh/FAQlist.html - - - *
* -  TIBS column archive -> http://www-lmmb.ncifcrf.gov/~pnh/readme.html - -  *
* - The BEST Molecular Biology HomePage -> http://www-lmmb.ncifcrf.gov/~pnh/  *
*******************************************************************************



More information about the Methods mailing list