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2 Questions about my agarose gel.

2 Questions about my agarose gel. - DNA Techniques

2 Questions about my agarose gel. - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


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  #1  
Old 12-06-2008, 11:23 PM
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Default 2 Questions about my agarose gel.



Okay so here is my gel.



Does anyone know some reasons why the bands in lanes 3,4,5 and 6 are gradually running slower and slanted?

Also, the first band seen on lane 3,4, and 5 are known to be "common", but does anyone know an explanation for why the person for lane 6 does not have it? Could it be that that person just doesn't have that particular gene sequence?

Thanks in advance!
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Old 12-08-2008, 05:40 PM
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Default Re: 2 Questions about my agarose gel.

for the first question. it is most likely due to the fact there are empty lanes in right side. I practice balancing sample loading in gel. example for gel above u could start loading at lane no. 2 instead of no. 1. this will give a equal voltage distribution (i hope my term is correct). just try it and see any improvement or not.

2nd question is a bit tricky, can u tell us what common gene is that? if it is high copy number, yes most probably subject for lane 6 does not have it. if not maybe extraction problem? too few data to make a proper suggestion. if you r running multiplex for sample in lane 6 u could try a singleplex and see how.
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Old 12-09-2008, 10:36 AM
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Default Re: 2 Questions about my agarose gel.

Have you tried doing another electrophoresis using the same electrophoretic set, voltage, buffer etc?
It looks like voltage distribution is uneven throughout the gel, maybe the device itself is a problem?
I don't think it's loading the samples - I also load my DNA samples starting from the left and don't always use all the lanes. But I recommend to load DNA mass marker also after the last sample lane (so 2 per gel).

Second thing - well, that may be a person not having this particular sequence, a rare example, but still possible. I don't know though, what 'common' sequence you have there, but I guess 'common' doesn't mean it has to be obligatory.
If you have doubts check it again.

Last edited by Aga; 12-09-2008 at 10:40 AM.
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Old 12-10-2008, 12:14 AM
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Default Re: 2 Questions about my agarose gel.

its for genotyping the ADH2 gene. Thank you all so much.
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