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#1 Arwa Husain

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Posted 22 April 2015 - 02:55 AM

Hi every1, I had been doing agarose gel electrophoresis and it has worked out so well every time. But since 3-4 days I have started facing problems with gel, it appears patchy on gel doc and also both the ends of gel get curved when it is put in buffer. Even the ladder is not running properly, which clearly means that there is nothing wrong with samples. Can someone please guide 



#2 Arwa Husain

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Posted 22 April 2015 - 02:56 AM

Hi every1, I had been doing agarose gel electrophoresis and it has worked out so well every time. But since 3-4 days I have started facing problems with gel, it appears patchy on gel doc and also both the ends of gel get curved when it is put in buffer. Even the ladder is not running properly, which clearly means that there is nothing wrong with samples. Can someone please guide 



#3 mdfenko

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Posted 22 April 2015 - 04:14 AM

can you show us a picture and give us more information about your procedure?

 

are you using anything new or newly prepared?

 

if so, did someone different prepare them this time?

 

how does the gel look to the naked eye? if it looks patchy then you probably have uneven melting and/or gelling and/or mixing of the agarose.

 

edges curling would indicate that the gel had dried somewhat and is rehydrating from the edges inward.


Edited by mdfenko, 22 April 2015 - 04:14 AM.

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#4 pavoni.ernesto

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Posted 22 April 2015 - 06:30 AM

Hi,

perhaps depends on the buffer (wrong concentration of salt or not deionized water)



#5 phage434

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Posted 22 April 2015 - 09:56 AM

Classic problems include making the gel with water instead of buffer, and inadequate heating of the gel, resulting in a non-uniform agarose solution.  The gel solutions should be crystal clear before being poured, and thoroughly mixed.  Pouring the gels while the solution is still hot can also cause problems.  Ideally, the crystal clear, well mixed solution should be cooled to around 60 before being poured.



#6 hobglobin

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Posted 22 April 2015 - 10:33 AM

I merged the posts. Please don't post double and then with a completely nondescript title (this will reduce input a lot).

Since here are many threads with electrophoresis problems, and you know how your gels look and what the experimetal details are (which you don't give for us), you are surely able to find the solution in the old threads. 


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