An example of DNA sequencing from a research based paper by me from Zhejiang Ocean University

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DNA sequencing

COI amplification used the pair of primers, FishF1 and FishR1(Ward et al., 2005). PCR was carried out in Thermo Cycler with a 25 μL volumes, containing 8.5 μL sterile nuclease free water, 2 μL of each primer , 12.5 μL Go Taq® Green Master Mix (Promega Inc, U.S.A.) and 2 μL of template DNA. PCR program used a thermal profile, including a preliminary denaturation for 2 minutes at 95 °C followed by 35 cycles of denaturation at 95 °C for 40 seconds, annealing at 52°C for 40 seconds, and extension at 72°C for 90 seconds, and finally a single extra extension at 72°C for 5 minutes. PCR products were confirmed via gel electrophoresis using 1.5% agarose gel stained with SybrGreen fluorescent dye for band characterization through Gel Imaging System. Subsequently, PCR products were sent for sequencing at Sangon Biotech Cooperation Ltd (Shanghai, P.R. China)

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