一种适用于梅花鹿源性产品鉴定的微量组织DNA提取方法的建立  

Establishment of a Micro-Tissue DNA Extraction Method Suitable for the Identification of Sika Deer-Derived Products

作　　者：任雨贺 金美伶[1] 宋超[1] REN Yuhe;JIN Meiling;SONG Chao(Institute of Special Animal and Plant Sciences of Chinese Academy of Agricultural Sciences,Changchun 130112,China)

机构地区：[1]中国农业科学院特产研究所,吉林长春130112

出　　处：《特产研究》2024年第5期54-57,共4页Special Wild Economic Animal and Plant Research

基　　金：吉林省科技发展计划项目(20220401113YY)。

摘　　要：为建立一种适宜梅花鹿源性产品分子鉴定的高效、便捷微量组织基因组DNA提取的方法。本研究分别以鲜品梅花鹿茸、风干梅花鹿茸、烘干梅花鹿茸、鲜品梅花鹿鞭、烘干梅花鹿鞭、风干梅花鹿鞭、鲜品梅花鹿肉、干品梅花鹿尾、梅花鹿血为样本,采用Chelex-100树脂溶液和蛋白酶K结合的方法提取基因组DNA,并检测其浓度与纯度;以此DNA为模板,扩增梅花鹿线粒体DNA的Dloop区特异性基因,并对PCR扩增产物进行琼脂糖核酸电泳及测序。结果表明,基于Chelex-100树脂提取法获得的基因组DNA模板浓度为52.57~63.39 ng/μL,A260/A280比值为1.75~1.96;梅花鹿线粒体特异性基因扩增产物电泳条带清晰,且长度准确;扩增产物测序,相似性大于95%。Chelex-100树脂提取法能够提取微量梅花鹿组织样本基因组DNA,且此方法低价、简便,适用于梅花鹿源性产品的真伪鉴定。In order to establish an efficient and convenient method for microtissue genomic DNA extraction suitable for molecular identification of sika deer-derived products.In this study,fresh sika deer antler velvet,air-dried sika antler velvet,dried sika antler velvet,fresh sika deer whip,dried sika deer whip,air-dried sika deer whip,fresh sika venison meat,dried sika deer tail and sika deer blood were used as samples,genomic DNAwas extracted by combining Chelex-100 resin solution with proteinase K,its concentration and purity were detected.Using this DNA as a template,the mitochondrial DNA D-loop region-specific gene of sika deer was amplified,the PCR amplification products were electrophoresis and sequenced.The results showed that the concentration of genomic DNAtemplate obtained by Chelex-100 resin extraction method was 52.57~63.39 ng/L.A260/A280 ratio is 1.75~1.96.The electrophoresis band of mitochondria-specific gene amplification products of sika deer is clear and the length is accurate;amplification product sequencing with similarity greater than 95%.Chelex-100 resin extraction method can extract the genomic DNAof small amount of sika deer tissue samples and this method is cheap,simple and suitable for the authenticity identification of sika deer origin products.

关 键 词：梅花鹿 Chelex-100树脂提取法 DNA提取 源性产品鉴定 

分 类 号：R282.5[医药卫生—中药学]