i have just joined this amazing forumo thanx for adding me to this forum.
i have a problem in my pcr reaction that is when i run a pcr i found non specific amplification. Master mix i use contain 5ul of buffer, MgCl2 1.8ul, DNTPse 0.6ul, Primers 1.5ul each F and R , Taq polymerase 0.5ul and water 18ul and 1.1ul of template.
thermal profile is 95c for 5minutes initial denaturation, 95 for 20sec, 59 for 30sec, 72 for 1 minute final extension 72c for 5minutes and holding temp. 4c.
what is your suggestion plz.