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Re-PCR on a gel purified PCR product resulted in smear. Why?


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#1 Curtis

Curtis

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Posted 27 July 2020 - 08:34 PM

I got a faint 4.3 kb pcr product on gel so I decided to purify and use it as template for a 2nd pcr with the same primers and machine program. Instead I ended up with a huge smear, although my own band of interest was visible at the top. I purified it but I want to know why I got a smear? was the annealing temp too low or the template was too much?




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