Digoxygenin End labelling

Klaus Salger salger at wap4.zi.biologie.uni-muenchen.de
Fri May 12 16:40:13 EST 1995


Wiggy (A.C.Hilton at bham.ac.uk) wrote:
: Hello,

: I am considering end-labelling a pBR322 hinf1 digest with DIG dUTP so I
: can get a marker for Southern blotting that I am doing.  I am concerned
: that the single (maximum 2) incorporated DIG-dUTP will be insufficient to
: give a detectable flourescent signal. Does anyone have any experience of
: end labelling with such non-isotopic methods at these low levels?

: I am not opposed to labelling with 35S or 32P but the development times on
: film are vastly different,  only 30 minutes with the DIG. 

: Are there any commercially available DIG markers?

: I already have a number of DIG labelled primers so a DIG marker would be
: great.

: Thnks for your time,

: Wiggy

Boehringer Mannheim sells DIG-labeled markers. Have a look into their
catalogue. I think they also label the markers by fill in of the digested DNA.
Detection should be no problem.

Cheers
  Klaus

(Standard disclaimer applies)

--
Klaus Salger                phone : ++49 (0)89 5902 -502
Zoologisches Institut       FAX   :                 -450
AG MacWilliams              e-mail: salger at zi.biologie.uni-muenchen.de
Luisenstr. 14
80333 Muenchen
Germany



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