摘要
目的优化深加工骨胶原蛋白肽粉中DNA的提取方法,鉴定检测其动物源性成分。方法采用2种试剂盒法,并对试剂盒部分步骤进行适度修改后提取骨胶原蛋白肽粉中的基因组DNA,分别用猪、羊引物和探针对提取的DNA片段进行实时荧光PCR扩增检测。结果只有DNeasy Blood&Tissue Kit提取试剂盒法提取的DNA纯度较好,A260/A280比值为1.73,浓度为8.2 ng/μL,并且经18S rDNA片段的PCR扩增结果表明样品中含有哺乳动物DNA,且DNA中不含抑制PCR反应物质。检测结果表明该骨胶原蛋白肽粉中含有猪源性成分。结论采用试剂盒法提取深加工骨胶原蛋白肽粉中DNA时,需对部分步骤做修改后才可进行。
Objective To optimize the method for extracting DNA from deep processed bone collagen peptide powder,and to identify and detect animal-derived components.Methods Totally 2 kit methods were used to extract genomic DNA from the collagen peptide powder after some steps of the kit were moderately modified.The extracted DNA fragments were amplified by real-time fluorescence PCR using primers and probes from pigs and sheep respectively.Results Only the DNA extracted by the DNeasy Blood&Tissue Kit extraction kit method had good purity.The A260/A280 ratio was 1.73 and the concentration was 8.2 ng/μL,and the PCR amplification results of the 18S rDNA fragment showed that the sample contained mammalian DNA,and the DNA did not contain a substance that inhibited the PCR reaction.Test results showed that the collagen peptide powder contained pig-derived ingredients.Conclusion When the kit method is used to extract DNA from deep-processed collagen peptide powder,some steps need to be modified before proceeding.
作者
沈晓丽
闫明俞
耿文宁
李海芳
SHEN Xiao-Li;YAN Ming-Yu;GENG Wen-Ning;LI Hai-Fang(Xinjiang Uygur Autonomous Region Institute for Drug Control,Urumqi 830054,China)
出处
《食品安全质量检测学报》
CAS
2020年第3期945-949,共5页
Journal of Food Safety and Quality
关键词
骨胶原蛋白肽
DNA提取
实时荧光PCR
源性成分检测
bone collagen peptide
DNA extraction
real-time fluorescence PCR
source component detection
作者简介
沈晓丽,硕士,实验师,主要研究方向为食品药品检验、分子生物学及药理学。E-mail:86513757@qq.com;通讯作者:李海芳,正高级实验师,主要研究方向为药品检验、药理毒理。E-mail:lhfhr@163.com
