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Old 01-22-2014, 11:46 AM   #2
Mike2188
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Location: Winnipeg

Join Date: Oct 2013
Posts: 26
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Hey,

I am surprised nobody has answered this yet, but it looks like you overamplified your library. What happens is that you run out of sequencing primers, whether as a result of a) not enough sequencing primer, b) too much template added, or c) running an excessive amount of PCR cycles. When the reactions run out of primer, the template starts annealing to itself and forming these long concatemers. You should try redoing the library with a smaller template input and fewer PCR cycles.

Hope this helps!
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