ICS 11.120.10 C23 团体标准 T/CACM 1027.104—2018 铁皮石斛 PCR鉴定 PCR identification of Dendrobii Officinalis Caulis 2018 - 11 - 19发布 2018 - 11 - 19实施 中 华 中 医 药 学 会 发布 T/CACM T/CACMT/CACM 1027.104 —2018 I 目 次 前言 ................................ ................................ ............... III 1 范围 ................................ ................................ .............. 1 2 规范性引用文件 ................................ ................................ .... 1 3 术语和定义 ................................ ................................ ........ 1 4 方案 ................................ ................................ .............. 2 5 仪器设备 ................................ ................................ .......... 2 5.1 离心机 ................................ ................................ ........ 2 5.2 金属浴 ................................ ................................ ........ 2 5.3 制冰机 ................................ ................................ ........ 2 5.4 涡旋振荡器 ................................ ................................ .... 2 5.5 超纯水发生器（分子生物学级别） ................................ ................ 2 5.6 高压灭菌锅 ................................ ................................ .... 2 5.7 PCR仪 ................................ ................................ ......... 2 5.8 电泳仪及其附件 ................................ ................................ 2 5.9 凝胶成像仪 ................................ ................................ .... 2 5.10 核酸蛋白仪 ................................ ................................ ... 2 5.11 电子天平（感量 0.01 g） ................................ ....................... 3 5.12 连续可调 微量移液枪 ................................ ........................... 3 5.13 低温冰箱 ................................ ................................ ..... 3 5.14 其他 ................................ ................................ ......... 3 6 试剂和溶液配制 ................................ ................................ .... 3 6.1 概述 ................................ ................................ .......... 3 6.2 三羟甲基氨基甲烷 -盐酸（Tris-HCl）溶液 ................................ ......... 3 6.3 0.5 mol/L 乙二胺四乙酸（ EDTA）溶液 ................................ ............ 3 6.4 前处理缓冲液 ................................ ................................ .. 3 6.5 CTAB提取缓冲液 ................................ ................................ 3 6.6 脱氧核糖核苷三磷酸 dNTP混合液 ................................ ................. 3 6.7 10× PCR缓冲液 ................................ ................................ . 3 6.8 不含3’→5’外切酶活性的 Taq酶（5 U/μL）................................ ......... 4 6.9 铁皮石斛检测用引物对序列 ................................ ...................... 4 6.10 引物溶液 ................................ ................................ ..... 4 6.11 琼脂糖（电泳纯） ................................ ............................. 4 6.12 DNA分子量标准 ................................ ................................ 4 T/CACM T/CACMT/CACM 1027.104 —2018 II 6.13 50× TAE电泳缓冲液 ................................ ............................ 4 6.14 6×上样缓冲液 ................................ ................................ . 4 7 检验程序 ................................ ................................ .......... 4 7.1 概述 ................................ ................................ .......... 4 7.2 取样 ................................ ................................ .......... 4 7.3 粉碎 ................................ ................................ .......... 5 7.4 DNA提取 ................................ ................................ ....... 5 7.5 PCR扩增 ................................ ................................ ....... 5 7.6 扩增产物检测 ................................ ................................ .. 6 8 结果判定 ................................ ................................ .........