So I am trying to amplify a ~13kb region of human DNA using LA-PCR. Currently I am using TaKaRa's kit and iProof by BioRad, but haven't been able to get a solid band on any of my gels. I was wondering if anyone else has had better luck with certain protocols or pcr programs/settings to get good amplification. Early on I was getting high smearing above 10kb. I've been messing with runtimes, adding betaine, and template concentrations but now I'm getting pretty much no amplification. Any comments would be appreciated, Thanks!
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