This is a gel with the trnH–psbA marker. Gradient PCR 54-57°C for a Nepeta species collected from nature (upper lanes) and a Centaurium species from in vitro conditions (lower lanes). We're confused by the presence of the two clear bands in Nepeta. Didn't sequence them. Can somebody suggest why this might have happened?
Branislav Šiler we have seen that pattern in some cases:
Article Design and evaluation of PCR primers for amplification of fo...
There may be some duplication in the cpDNA of that species. if available, check the chloroplast genome.
The primers may be binding in another region - you can try to increase the annealing temperature a few more degrees. there is no visible differences in the gradient you did, try a more drastic gradient to see if the second band disaperars.
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