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Wavy Lines in Gel

wavy lines gel electrophoresis

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#1 tihong10

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Posted 06 August 2013 - 05:19 AM

Hi,

 

When I run my pcr product (recombinant plasmid as template with insert primers) in 2% agarose gel (settings: 90V for 70min) I get these wavy lines. These lines I suspect are due to an incorrectly made gel, but I have searched for and tried many gel making protocols and still get these wavy bands; so, I am beginning to think maybe the product itself is somehow leading to this appearance.

 

However, I have rerun the product that initially resulted in wavy lines in gel lanes that have been run with positive control products and show straight bands, and sometimes get straight lines and sometimes get wavy lines, which leads me to believe that it indeed isn't the product itself but the gel or perhaps the settings that are leading to the wavy bands.

 

Can someone please let me know what this phenomenon is the result of?

 

Thank You

Ted

 

P.S. image attached

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  • Wavy Gel.jpg

Edited by tihong10, 06 August 2013 - 05:24 AM.


#2 phage434

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Posted 06 August 2013 - 06:03 AM

Some potential problems:

* Make sure your agarose is completely dissolved in buffer. The solution should be crystal clear. Swirl it to make sure it is completely mixed. (most common problem)

* Cool your agarose before pouring the gel. 70 would be a good target temperature, but definitely above 55.

* Allow the gel to completely set. Using gels before they set can cause problems.

* Remove the comb carefully. Excess gel in the well can cause distortions of this kind.



#3 tihong10

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Posted 06 August 2013 - 06:08 AM

Thanks Phage434,

 

I'll try these tips and see if the lines disappear.

 

-Ted







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