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non specific amplification in pcr

non specific amplification in pcr - Biochemistry Forum

non specific amplification in pcr - Discuss and post questions regarding the study of Biochemistry. If you need homework help this is the place to ask!


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Old 05-05-2013, 08:45 PM
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Default non specific amplification in pcr



sir!
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.
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