gradient pcr trouble
hi.. i am facing troble with gradiant pcr. its siRNA viral construct of about 900 bps. i have pcr it on 55-65 many times it shows a light band on 63 but nothing at 64C and 62C. at 63 it also shows a couple of non-specific DNA bands..i ve changed taq, quantity of taq, pcr buffer (kcl & NH4) both tested, dNTPs and primers working fine. any solution????
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