论文部分内容阅读
目的研制川贝母DNA检测试剂盒,确定试剂盒组成及操作程序,建立一种简便、快速鉴定川贝母的分子生物学检验方法。方法分别采用药典法(2010年版《中国药典》增补本)和试剂盒法提取川贝母基因组DNA,进行PCR反应和RFLP鉴定。结果采用药典法提取出的川贝母基因组DNA的OD260/OD280值最高为(1.57±0.05),而采用试剂盒法所提DNA的OD260/OD280值可达(1.73±0.10);两法PCR结果均在300 bp上方有单一明亮条带;RFLP鉴定图谱均在100~250 bp之间出现2条清晰条带。结论试剂盒法较药典法更稳定,且核酸提取量及纯度都高于药典法,2种方法 PCR及RFLP鉴定结果完全一致。川贝母DNA检测试剂盒特异性好、灵敏度高、稳定性强,适用于川贝母的快速检测。
Objective To develop the Fritillaria cirrhosa DNA Assay Kit, to determine the kit composition and operation procedure, to establish a simple and rapid identification of Fritillaria cirrhosa molecular biology test methods. Methods The genomic DNA of Fritillaria cirrhosa was extracted by the method of Pharmacopoeia (2010 edition of “Chinese Pharmacopoeia” Supplement) and kit method, and the PCR reaction and RFLP identification were carried out. Results The OD260 / OD280 value of the Fritillaria cirrhosa DNA extracted by the pharmacopoeia method was the highest (1.57 ± 0.05), while the OD260 / OD280 value of the DNA extracted by the kit method was (1.73 ± 0.10) All had a single bright band above 300 bp. Two clear bands appeared between 100 and 250 bp in RFLP identification. Conclusion The kit method is more stable than the pharmacopoeia method, and the amount of nucleic acid extracted and purity are higher than that of the pharmacopoeia method. PCR and RFLP identification results of the two methods are completely consistent. Fritillaria cirrhosa DNA testing kit specificity, sensitivity, stability, for Fritillaria rapid detection.