穿山甲鳞片荧光PCR鉴定方法研究

北京理工大学珠海学院2016级本科生毕业设计Study on the identification of pangolin scales by fluorescent PCR.AbstractAs the most serious species in the world,pangolin is becoming extinct.Because it is said that pangolinscales can cure many diseases,people are flocking to kill a large number of pangolins,and a large numberof illegally traded pangolin scales are circulating in the market.In this experiment,compared with the traditional method of identification of pangolin,theidentification effect is more accurate and the identification speed is faster.In this experiment,six groups ofDNA samples of pangolin were used as the experimental objects,and six groups of non-specific sampleswere used as the control group.The plasmid of pangolin was diluted in gradient,and 104 of them wereselected as positive.Different from the traditional morphological observation of pangolin scales,chromatograph detection and common PCR,fluorescent PCR has higher sensitivity and specificity,and itsfast and efficient characteristics provide more reliable backing for the results.Four groups of experiments were carried out in this experiment,including laboratory sample detection,specific detection,sensitivity detection and repeatability detection.Parallel tests will be set up for eachgroup.In sensitivity test,fluorescent PCR can capture DNA with dilution ratio of 10-8,which is twice thesensitivity of ordinary PCR.In repeatability,the standard deviation and coefficient of variation arecalculated according to CT value,and the results are all within 5%,which is relatively stable Amplification.The results showed that fluorescent PCR had a considerable advantage in the determination of DNAand scales of pangolin.Key words:real time fluorescence quantitative PCR;pangolin scales