DNA stuck in wells!

SerAln SerAln at yahoo.com
Fri Oct 25 08:06:16 EST 2002


I had the same problem when adding RNAse to the digestions. Don't know why, 
since we used RNAse in DNA restrictions for years without any problem. We first 
blamed on the supplier, so they gave us new vials.... no luck, the DNA always 
suffered a shift, no matter the way we prepared the RNAse. Now we add very 
diluted RNAse or no RNAse at all, and we add the RNAse during the DNA 
preparation prior phenolization step.

Hope this helps.

Sergio

Craig Lewis wrote:
> Yes, the dye moves normally. The strangest thing is that the problem wells
> occur quite randomly - not all gels and not all wells.
> 
> Kyle Legate wrote:
> 
> 
>>On Thu, 10 Oct 2002, Craig Lewis wrote:
>>
>>
>>>Has anyone experienced DNA from a simple restriction digest fail to move
>>>in an agarose gel? This is a relatively recent problem I'm having. It
>>>seems that the DNA gets stuck in the well. The gel is made with TAE
>>>buffer and good quality agarose with EtBr added to the gel. The DNA is
>>>in the 1 to 10 kb range and is quite clean. I'm using a
>>>sucrose/bromophenol blue loading dye.
>>>
>>
>>Does the dye move?
>>
>>... . . .  .  .  .    .    .    .     .     .     .      .      .      .
>>legatek at mcmaster.ca             Kyle Legate            legatek at hotmail.com
>>
>>   Tower of Tongues:Thursday PM:10:30-11:30 EDT:http://cfmu.mcmaster.ca
>>    moon musick:ritual:IDM:experimental(electronica):minimalism:glitch
>>.      .      .     .     .     .    .    .    .   .   .   .  .  . . ...
> 
> 




More information about the Methods mailing list