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AFLP smear

AFLP smear - Protocols and Methods Forum

AFLP smear - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

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Old 08-13-2009, 10:12 AM
Pipette Filler
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Default AFLP smear

I am working with AFLP,s in elasmobrachs, and at the beginning I get pretty good result, however, after a few samples working well, now I can not get any peak in my chromatogram, I have check the quality of DNA, very good, I have check that the digestion/ligation working well, and even my preselect amplification produce a great smear between 100 and 1500 pb. But when I run final PCR product in an agarose gel, I get as bright smear in from 500 to the tank (the hole when you put the DNA), and of course I donít get nothing in the chromatogram.
I have changed concentration of MgCl and DNA, I have changed the number of cycles in the PCR profile, I have changed all my chemicals, and nothing has worked.
So, any idea of what could produce the smear at large number of pb?
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