《Table 1.List of primer used for the amplification of the four pierid butterfly mitogenome.》

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《Mitochondrial genomes of four pierid butterfly species(Lepidoptera: Pieridae) with assessments about Pieridae phylogeny upon multiple mitogenomic datasets》

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*Annealing temperature

Total genomic DNA was extracted from leg muscle of each adult individual by the glass powder method with minor modifications（Hao et al.，2005）.Three partial fragments of COI，ND4，and Cytb were amplified by using standard short primers from Simon et al.（1994），other primers for the amplications of long and short fragments were designed by Primer Premier 5.0（Singh et al.，1998）through multiple sequence alignments of butterfly mitogemomes using Clustal X1.8（Thompson et al.，1997）.All the primers were synthesized by Sangon Biotechnology Co.Ltd.（Shanghai，China）（Table 1） .Long PCR amplification reactions were performed under the following conditions:an initial denaturation at 95°C for 5 min；30 cycles of denaturation（denaturing at 95°C for 50 s，annealing at 50°–55°C for 50 s，extension at 68°C for 150 s），and a final extension step at 68°C for 10 min.The PCR products were separated by 1.2%agarose gel electrophoresis，purified by using a 3S Spin PCR Product Purification Kit（Sangon Biotechnology Co.Ltd.，Shanghai）and sequenced on an ABI-377automatic DNA sequences.For each long PCR product，the complete，double-stranded sequence was determined by the“primer walking”method.