限制性複製片段長度多型在動物種屬鑑定之研究

• Main Author: 洪聖儀
• Other Authors: 李俊億
• Format: Others
• Language: zh-TW
• Published: 1998
• Online Access: http://ndltd.ncl.edu.tw/handle/q92chs

碩士 === 中央警察大學 === 鑑識科學研究所 === 86 === 　　Amplified Fragment Length Polymorphisms (AFLP) was developed by Zabeau and Vos in 1993 which can produce a novel DNA profiles. This method was a DNA fingerprinting technique that combined RFLP and PCR to amplify specific restriction fragments. Typically 30-100 bands are amplified and separated on denaturing polyacrylamide gel. It can be used for DNA identification on any biological origin with minute specimen. In this study we collected the tissue samples of 13 different animals (including seven species of endangered animals in Taiwan and six species of livestocks) and used AFLP technique to analyze and establish an AFLP procedure of species identification. There was no radioactive isotope applied in this study, but visualized the whole AFLP patterns by silver stain method. With the radioactive AFLP procedure, only one the labeled fragments were visualized on the autoradiographic films, and AFLP patterns were much more simple. In this study we use silver stain method to present all the amplified fragments, and it can avoid the disadvantages of tedious and hazardous process to health. Besides, we also used automated gel stainer to stain the gel and it took only two hours. 　　This study used 29 primer pair can be used on animal species. According to the AFLP patterns, the primer pairs B2, B3, B4 can get optimum results, and can also apply to differentiate livestock samples successfully. The AFLP patterns produced by these primer pairs have obvious diversity among different animal species. Finally, we used UPGMA method to proceed cluster analysis and draw the dendrogram. We discovered that even the same species of animals can be clustered together by single primer pair''s AFLP patterns, but it was still not enough to determine the species. On the other hand, we find DNA quality will influence the AFLP result, so it is important to collect fresh sample to get specific AFLP fingerpring. This study had accumulated all AFLP patterns using 29 primer pairs to establish a databank. The databank will provide helpful references on animal species identification.