artifacts in sequencing gels

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artifacts in sequencing gels

Postby ng1 » Aug 19 2017 4:50 am

Hi everyone,

I am using sequencing gels to separate radiolabeled (32P dATP) DNA fragments that are generated during DNA repair. I often encounter smear-like artifacts in empty lanes (that are not loaded with any samples) close to loaded lanes. This prevents me from being able to interpret the data. Does anyone know the cause of this kind of artifact and how It can be prevented?

I run 4 mm thick 7% polyacrylamade gels using the S2 sequencing gel electrophoresis apparatus and shark tooth combs.
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Re: artifacts in sequencing gels

Postby mdfenko » Aug 21 2017 12:10 pm

a picture would help...

smears like that usually come from sample bleeding through the gel, crossing the comb's tooth and separation of the gel from the plate (eg air pockets).
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Re: artifacts in sequencing gels

Postby relaxin » Aug 24 2017 12:11 am

You can prevent this horizontal leaking by using comb that has square teeth and freshly prepared gel (old gel may shrink).
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