摘要
目的本文以阿胶传统工艺加工过程样品为研究对象,探讨关键加工阶段阿胶样品中驴基因组DNA核基因和线粒体基因的降解变化规律。方法提取阿胶工艺过程中的驴基因组DNA,分别利用超微量分光光度计、琼脂糖凝胶电泳、普通PCR、荧光定量PCR和简单序列重复（SSR）微卫星毛细管电泳（CE）检测方法对提取的DNA质量进行评价。结果研究结果显示,在原料驴皮、化皮、双效、浓缩和加辅料阶段,琼脂糖凝胶电泳均能检测到清晰的条带,普通PCR可扩增到200~1 600 bp左右的片段,而在凝胶阶段DNA降解最为严重,只能利用琼脂糖凝胶电泳检测到100~800 bp的目的片段,利用荧光定量PCR检测技术线粒体基因在阿胶加工各个阶段均可检测出,核基因凝胶阶段未检出;通过PCR-SSR-CE检测在凝胶阶段核基因未检出特征峰,更进一步证明了凝胶阶段DNA核基因降解严重。结论对阿胶工艺加工过程DNA变化规律发现,随着阿胶加工的深入进行,各个阶段DNA发生不同程度的降解,其中凝胶成品阶段降解最为严重,本研究对比普通PCR、荧光定量PCR和简单序列重复微卫星毛细管电泳检测方法,发现荧光定量PCR检测方法最快速、灵敏和可靠,通过比较发现以线粒体基因为靶基因在阿胶的各个加工阶段均能满足DNA溯源的要求。
Objective In this paper,each key processing stage samples of Colla Corii Asini as the research object to explore for DNA degradation of process variation in Colla Corii Asini. Methods Extractive DNA was evaluated by ultra trace spectrophotometric meter detection,agarose gel electrophoresis,PCR,fluorogenic quantitative PCR and SSR microsatellite CE detection.Results The results showed that in raw materials,melting stage,dual processing stage,before the condensed phase and adding accessories stage can detect clear amplified bands in the size range in the 200 ~ 1 600 bp fragment.The degradation of DNA was the most serious in the sample of the gel paste,and the range of 100 ~ 800 bp could only be detected.Conclusion With the processing of Colla Corii Asini product,DNA quality was gradually declined,and that DNA degraded most seriously at the glue paste stage,but the genomic DNA was still competent for Colla Corii Asini materials identification and DNA traceability.
出处
《药学研究》
CAS
2016年第9期501-507,共7页
Journal of Pharmaceutical Research
基金
山东大学高校院所自主创新项目(No.201401264)
关键词
驴皮
阿胶
DNA降解
质量检测
分子生物学
Donkey hide
Colla Corii Asini
DNA degradation
Quality detection
Molecular biological
