摘要
目的研究肉类线粒体DNA(mitochondrial DNA,mtDNA)的提取方法,对8种生肉中貂源性成分进行检测,建立生肉中貂源性成分PCR检测方法.方法应用改良SDS碱变性法和盐析法提取肉类mtDNA,设计貂肉细胞色素b(cytochromeb,cytb)特异性引物,应用PCR对8种生肉中貂源性成分进行鉴别.结果两种方法提取肉类mtDNA的浓度和纯度均能保证PCR要求,改良SDS碱变性法提取的貂DNA样品纯度和产率较高;貂引物可以从8种生肉中鉴别出貂源性成分;经过多次重复试验证明了貂肉引物的特异性.结论该方法快速简便,为肉制品质量监控提供了实验基础.
Objective To explore a practical method for extraction of mitochondrial DNA from Martes zibellina meat and to set up a method to distinguish the Martes zibellina from raw meat by PCR technology.Method The improved SDS alkaline lysis and salt-outing methods were used to extract mtDNA from Martes zibellina meat.The specific primers of Martes zibellina cytochromeb(cytb)was designed to identify the most common adulterated meat.Results With achieving PCR response requirements,the mtDNA from Martes zibellina meat with content and purity can be got with two kinds of methods.The content and purity of improved SDS alkaline lysis method was higher than that of salt-outing method.Martes zibellina source composition can be identified from 8 kinds of raw meat by Martes zibellina primer,the specificity of Martes zibellina primers was proved through the repeating experiment.Conclusion Extraction of mtDNA by improved SDS alkaline lysis method is more practical and accurate,which can provide experimental basis of application for the meat products quality monitoring.
作者
艾金霞
周童
李明成
孙丽媛
高丽君
夏薇
苑广信
张丽华
AI Jinxia;ZHOU Tong;LI Mingcheng;SUN Liyuan;GAO Lijun;XIA Wei;YUAN Guangxin;ZHANG Lihua(Medical Technology College of Beihua University,Jilin 132013,China;Pharmacy College of Beihua University,Jilin 132013,China;Leibo Science and Technology Ltd,Jilin 132013,China)
出处
《北华大学学报（自然科学版）》
CAS
2022年第6期789-793,共5页
Journal of Beihua University（Natural Science）
基金
吉林省科技发展计划项目(20200404152YY,20210204185YY)
北华大学医学技术学院“晨裕”科研创新项目(BH007).
关键词
改良SDS碱变性法
盐析法
貂源性成分
线粒体DNA
PCR
improved SDS alkaline lysis method
salt-outing method
Martes zibellina source composition
mitochondrial DNA
PCR
