A study was conducted on the identifications of the degraded samples of sika deer (Cervus nippon) and red deer (Cervus elaphus) by phylogenetic and nucleotide distance analysis of partial Cytb and 12s rRNA genes s...A study was conducted on the identifications of the degraded samples of sika deer (Cervus nippon) and red deer (Cervus elaphus) by phylogenetic and nucleotide distance analysis of partial Cytb and 12s rRNA genes sequences. 402 bp Cytb genes were achieved by PCR-sequencing using DNA extracted from 8 case samples, and contrasted with 27 sequences of Cytb gene downloaded from GenBank database. The values of three nucleotide distance between three suspected samples and sika deer were identical (0.026±0.006), which was smaller than the smallest nucleotide distance between eastern red deer and sika deer (0.036). Furthermore, phylogenetic analysis of sika deer and red deer indicated that the evidences located within the same cluster as sika deer. The evidences were sika deer materials. As the same way, other three suspected samples were derived from red deer. The results were further confirmed by phylogenetic and nucleotide distance analysis of 387 bp 12s rRNA gene. The method was powerful and less time-consuming and helpful to reduce the related cases with wildlife.展开更多
[Objective] To introduce an improved method for DNA extraction from the faeces of red deer. [Method] Based on the traditional method of CTAB lysis, we proposed an improved DNA extraction method according to the charac...[Objective] To introduce an improved method for DNA extraction from the faeces of red deer. [Method] Based on the traditional method of CTAB lysis, we proposed an improved DNA extraction method according to the characteristics of red deer faeces. [Result] This improved method extracted high-quality fecal DNA from Tianshan red deer and amplified the mitochondrial cytochrome b gene. With the muscle and fur DNA of red deer as the control, the sequencing results further con- firmed the reliability of the method. [Conclusion] The method requires no proteinase K in the process of extraction, and the extracted DNA can be used for PCR ampli- fication directly without the purification of DNA purification kit, thus, it is cost-saving.展开更多
文摘A study was conducted on the identifications of the degraded samples of sika deer (Cervus nippon) and red deer (Cervus elaphus) by phylogenetic and nucleotide distance analysis of partial Cytb and 12s rRNA genes sequences. 402 bp Cytb genes were achieved by PCR-sequencing using DNA extracted from 8 case samples, and contrasted with 27 sequences of Cytb gene downloaded from GenBank database. The values of three nucleotide distance between three suspected samples and sika deer were identical (0.026±0.006), which was smaller than the smallest nucleotide distance between eastern red deer and sika deer (0.036). Furthermore, phylogenetic analysis of sika deer and red deer indicated that the evidences located within the same cluster as sika deer. The evidences were sika deer materials. As the same way, other three suspected samples were derived from red deer. The results were further confirmed by phylogenetic and nucleotide distance analysis of 387 bp 12s rRNA gene. The method was powerful and less time-consuming and helpful to reduce the related cases with wildlife.
基金Supported by the National Natural Science Foundation of China(31060152)the Natural Science Foundation of the Xinjiang Uygur Autonomous Region,China(2010211A02)the Key Program for Animal Sciences of Xinjiang University,China~~
文摘[Objective] To introduce an improved method for DNA extraction from the faeces of red deer. [Method] Based on the traditional method of CTAB lysis, we proposed an improved DNA extraction method according to the characteristics of red deer faeces. [Result] This improved method extracted high-quality fecal DNA from Tianshan red deer and amplified the mitochondrial cytochrome b gene. With the muscle and fur DNA of red deer as the control, the sequencing results further con- firmed the reliability of the method. [Conclusion] The method requires no proteinase K in the process of extraction, and the extracted DNA can be used for PCR ampli- fication directly without the purification of DNA purification kit, thus, it is cost-saving.