Gel purification ligation problem

Wolfgang Schechinger wolfsc at ibms.sinica.edu.tw
Thu Jul 19 03:10:01 EST 2001




> For the UV box thing: a plain blue light box lights up EtBr, you
> then need orange filter to see it (blue and orange
> plexiglass/plastic). There's one commercially available, can't
> remember the company though.
> 

named 'dark reader', however the signal is not a strong as with 
UV (but for cloning it should be ok). 

Wolfgang
> cheers
> 
> Bruno
> 
> 
> "Dr. Harry Witchel" <Harry.Witchel at deletethis.bristol.ac.uk>
> wrote in message
> news:3B558261.8F6053E9 at deletethis.bristol.ac.uk... > Hi folks --
> > In my lab we are currently having problems with gel >
> purification of DNA from agarose.  We are using Qiagen gel
> purification > kits with all of the extra steps, and we are
> getting good yields of > fragments (as judged by running an
> aliquot of the yield on a gel), but > absolutely everything we
> purify is unligatable (as judged by failed >
> ligation-transformations).  We know the restriction enzymes (and
> ligase > and competent bugs) are working because if we do not gel
> purify, we can > get > the vectors to circularise (ie non-gel
> purified vector + no ligase --> > 14 colonies, while non-gel
> purified vector + ligase --> 200 colonies). > We suspect there
> may be something coming out of our gel purification > step that
> contaminates ligation; we currently purify from Biorad >
> molecular biology grade normal melting point agarose.  Do you
> have any > suggestions or ways to move forward on the gel
> purification front?  When > you reply, please note that you have
> to delete "deletethis" from my > email address. > All the best, >
> Harry > > -- > > Dr. Harry J. Witchel > Dept. Physiology >
> Medical School > Bristol  BS8 1TD >    England > 44-(0)177-928
> 7817 > Harry.Witchel at deletethis.Bristol.ac.uk > To send me an
> email, use the above address, but delete "deletethis." > from the
> address > http://www.bris.ac.uk/Depts/Physiology/Staff/HW/hw.htm
> 
> 
> 


[tiss mezzage wahs broduceRd using TYPO GENERATOR zoffwer]
-----
Dr. Wolfgang Schechinger
Lab N233 (c/o Dr. Steve Roffler)
Institute of Biomedical Sciences
Academia Sinica
128 Yen-Chio Yuan Rd. Sec.2
Taipei 115
Taiwan R.O.C.
Tel +886-2-2789-9152
Fax +886-2-2782-9142
Mobile +886-925-136893
e mail wolfsc at ibms dot sinica dot edu dot tw
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