; Molecular Biology
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> From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of Opher
> Gileadi [opher.gile...@sgc.ox.ac.uk]
> Sent: Saturday, September 30, 2017 3:44 PM
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Subject: Re: [ccp4bb] Off topic: denaturing urea gels
In addition to the previous suggestions:
With very small gels, the sample composition and depth (in the well) have a
strong effect on the resolution.
Rinse the wells with TBE buffer just before loading, as urea from the gel
diff
In addition to the previous suggestions:
With very small gels, the sample composition and depth (in the well) have a
strong effect on the resolution.
Rinse the wells with TBE buffer just before loading, as urea from the gel
diffuses into the well and may prevent the sample from settling at the
Smith:
One should expect to see ladders each separated by 1 nt in such cases.
Mohammed:
My suggestions:
1) running at 70V seems low. I do not see a reason for not using higher
voltages. 200Vx45min should be OK. IT is better to not let the BPB front run
out of the gel - so that you know whether
your enzyme cannot give definite fragment. thus smear
发自网易邮箱大师
在2017年09月30日 19:28，Mohammad Khan 写道：
Dear all,
I am working with an exonuclease and I run the digested DNA on a
8Murea-20%acrylamide gel in TBE buffer. I use the Mini-Protean BioRad system
and cast gels of about 8.6x6.5 cm
Dear all,
I am working with an exonuclease and I run the digested DNA on a
8Murea-20%acrylamide gel in TBE buffer. I use the Mini-Protean BioRad
system and cast gels of about 8.6x6.5 cm dimensions with 1.5 mm thickness.
I use a 15 well comb. I run my gels at 70 V for as long as 4 hours till my
