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In nested PCR not sharp bands
i'm doing nested PCR of microscopically positive stool samples. but after primary PCR there are no visible bands on gel and in nested pcr bands which r there r light. further i have to for restriction digestion then how can proceed with these light bands or what should i do for increasing the sharpness of my bands.
Re: In nested PCR not sharp bands
You either need more DNA template at beginning of PCR, more cycles --or DNA cleanup before PCR; stool can have PCR inhibitors, easy fix is running samples through a Chromaspin column.
|bands , nested , pcr , sharp|
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