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Syber green gel staining after electrophoresis


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#1 Tatiana Dimitrieva

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Posted 26 March 2015 - 02:04 AM

Hi there,

Please help me solve the problem with Syber green handeling, I use 1x solution to stain gel after electrophoresis and get smearing bands in DNA ladder (It's ok in the case of adding Syber green directly during the loading).  How to avoid it? I tried to vary loading amount, voltage, time without any differences.

 

Tatiana Dimitrieva

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Edited by Tatiana Dimitrieva, 26 March 2015 - 03:03 AM.


#2 Trof

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Posted 26 March 2015 - 02:29 AM

Which SYBR green product you have? I have no good experience with post-run staining in general. The dye only slowly penetrates inside the gel, often binding more unspecific smears. I put the dye into the gel instead before casting, whether it is EtBr or alternative. I've seen this working well with this SYBR.

 

But if your samples look fine and only the ladder is smeared, did you check the ladder is fine?


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#3 Tatiana Dimitrieva

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Posted 26 March 2015 - 02:38 AM

Thank you for the answer, 

I use the same Sybr as you,

and ladder is fine, I always use it with EtBr without any problems.



#4 Trof

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Posted 26 March 2015 - 02:48 AM

I would consider if adding the SYBR directly into the gel isn't a better solution (but it will leave traces in the buffer and in the tank, you either need to have a separate tank for each dye or change the buffer and clean the tank between uses, but that is not a huge problem, we are currently using both EtBr and alternative dye in the same set of tanks).


Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

'Normal' is a dryer setting. - Elizabeth Moon


#5 Tatiana Dimitrieva

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Posted 26 March 2015 - 04:14 AM

Ok. Thanks for the concern.






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