Non radioactive

[Dr. Klaus Eimert]

James Abbott wrote:
> 
> curtis alexander wrote:
> 
> We have tried a couple of different kits, generally fairly > succesfully. For Westerns, both ECL kits from Amersham and a kit from > NEB seem successful, the NEB kit produced fewer artifacts and less 
> background than ECL, but Amersham have since produced a new version of > the kit which is meant to be better. 
> For Southerns and colony blots, we have used ECL again, although this > hasn't been as good for low target concentrations. In one case we also > tried Amershams Gene Imager kit for use in low target applications but > without success. 
> By far the best results have come using DIG labelling from Boehringer
> Mannheim, although using the kit as directed did not produce 
> particularly impressive results. Following the protocol of Blum et al > (1993) (see below for reference) produced startlingly good results. 
> The blot used with Amershams Gene Imager was stripped, and reprobed > with a DIG labelled riboprobe and this protocol and produced bands we > had not been able to see using 32P - very impressive stuff, and 
> completely safe. 
> The reference for the protocol is below: 
> Authors - Blum G.E., Meier, M., Frank, J., Muller, G.A.
> Year - 1993
> Title - Reduction of Background Problems in Nonradioactive Northern > and Southern Blot Analysys Enables Higher Sensitivity Then 32P-Based
> Hybridizations
> Journal - Anal. Biochem
> Volume - 210
> Pages - 235-244
> 
> Hope this is of some use.
> Happy Blotting,
> James





Hi,

sounds interesting! Since I have some trouble to get the kit working
well I would be interested in trying the mentioned protocol. However, 
we don't have access to Anal. Biochem. in our area :-(
Could anybody using the method post a short protocol, please? I would
really appreciate!




Klaus


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 Dr. Klaus Eimert
 Dept. of Botany
 Geisenheim Research Institute		 
 Von-Lade-Str. 1                            Phone: + 49 6722 502 469
 D-65366 Geisenheim                         FAX  : + 49 6722 502 460
 Germany                E-mail: eimert at geisenheim.fa.fh-wiesbaden.de
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