AflII restriction enzyme

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	The amplified fragments were digested with restriction endonucleases (ApaI, AflII, BglI, or HpaII; Takara Biomedical) and separated on 8% polyacrylamide gels, which were stained with SYBR Green I nucleic acid stain (Takara Biomedical).

Protocol tips
The amplified fragments were digested with restriction endonucleases (ApaI, AflII, BglI, or HpaII; Takara Biomedical) and separated on 8% polyacrylamide gels, which were stained with SYBR Green I nucleic acid stain (Takara Biomedical).

Publication protocol

All regions of the mitochondrial genome were amplified by PCR with the GeneAmp PCR 9600 System (Perkin-Elmer, Norwalk, CT, USA). PCR amplifications were performed with 200 μM of dNTPs, 0.5 μM of forward and reverse primers, 1 U of ExTaq polymerase (Takara Biomedical, Ohtsu, Japan), 5 μl of 10× PCR reaction buffer (Takara Biomedical), and 200 ng of DNA template, for a total of 50 μl. The primers used in this study are shown in [Table 1]. The amplification consisted of an initial 1 min at 94°C, followed by 35 cycles of 94°C for 30 s, 53-62°C for 60 s, and 72°C for 10 min. The amplified fragments were digested with restriction endonucleases (ApaI, AflII, BglI, or HpaII; Takara Biomedical) and separated on 8% polyacrylamide gels, which were stained with SYBR Green I nucleic acid stain (Takara Biomedical).

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Manufacturer protocol

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