Rhodamine dUTP and capillary eletrophoresis

Edward Weiss via methods%40net.bio.net (by edward.weiss from utoronto.ca)
Wed Dec 12 15:50:55 EST 2007


I don't think the salt concentration is a particular issue -- we've done 
exactly the same procedure using labeled primers instead of labeled 
dUTP, and it works fine. I don't imagine the salt concentration of the 
dUTP stock would be so high as to make things significantly different in 
the final product. I've tried 1/10 and 1/20 dilutions too, with no luck.

Thanks for the help,

Ed

chovek69 wrote:
> Yeah Wolfgang makes a good point. Capillary electrophoresis is very
> sensitive to salt concentrations above the regular PCR ones (e.g in
> restrictase buffers). Try desalting or diluting samples 1/5 or 1/10
> with ddH2O before separation.
>
> Hope this will help
>
> On Dec 12, 10:07 am, WS <novalidaddr... from nurfuerspam.de> wrote:
>   
>> Dear Ed,
>>
>> have you tried desalting or doing a (PCR) cleanup before you analyze
>> the samples?
>>
>> Best regards,
>>
>> Wolfgang
>>     
>
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-- 
Edward S. Weiss
Research Technologist
Genetic Analysis Facility
The Centre for Applied Genomics @
The Hospital for Sick Children
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101 College Street
Toronto, ON M5G 1L7
Canada

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edward.weiss from utoronto.ca



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