Filter combs & loading protocol (Use of Ficoll)
jarmo.korkko at drexel.edu
Tue Nov 30 05:42:54 EST 1999
We have a slight problem with the Ficoll- protocol with filter combs (The
The water loading protol actually works quite well - some overlapping of the
samples but they are still trackable/readable.
I tried to load with 20% Ficoll protocol (add Ficoll to "loading area",
insert the comb with samples, add 1xTBE buffer), run for 30sec-1min, rinse
Ficoll out, remove the comb - RUN). Results:
1. Relatively low peak height in first 100bp
2. Closer to 100bp peaks started to get wider
3. Around 100bp ~4bp compression and after that the nicest sequence ever all
the way to 800 or so bp
4. Beatifull lane separation even though all the samples were loaded at the
Same happened with 15% Ficoll...
Has anyone experienced similar problem? It reminds me a little bit of urea
effect or glycerol effect? I haven't tried with taurine buffer yet so I'm
not sure if that would help.
If we can get this working I'm not even considering alternative loading
methods...it's very easy and the lane separation was unbelievable...
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