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Table 4. Preparation of Primer and gDNA template Final reaction volume for one PCR is 25μl Reaction In this table you are calculating volumes for 3.5 reactions Stock concentrations components Master mix 1 Master mix 2 Final conc. or μl to add Final conc. or μl to add amount amount Primer Concentration* 2μM 0.2μM 0.4μM (μm) Amount of gDNA 5ng/μl 25ng 25ng template (ng) One Taq 2X 1X 1X Mastermix Nuclease-free Add water such that the final total reaction volume in each tube is 3.5 x 25μl water * Forward and reverse primers are prepared in a single tube: a stock concentration of 2μM means that the solution contains the forward and reverse primers at 2μM each