Hello,
What technique or modification to the gel electrophoresis procedure do I need in order to increase the difference in distance travelled by two alleles with very similar sizes? In my gel electrophoresis result (attached), the A. texanum samples' bands are very close to the A. jeffersonianum's. How do I magnify this differences in order to clearly distinguish between the two alleles from these two different species of salamanders?
Extract the fractionated similar size bp sequences and multiply the one u want with PCR by adding the primers corresponding to the sequence.This will increase the quantiy of one allele and then you can discriminate based on the quantity in gel electrophores @ Minh Le
It can might be done by tagging a fluoroscence dye to one of the bp to distinguish among the two similar size@ Minh Le
You can increase the concentration of agarose in your gel--maybe try 3% (assuming your bands are 1-200bp in the image).
You could also try running a polyacrylamide gel--these are optimal for resolving shorter amplicons (see https://www.idtdna.com/pages/education/decoded/article/running-agarose-and-polyacrylamide-gels)
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