常用中药的生物鉴定课件

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,*,常用中药的生物鉴定,常用中药的生物鉴定方法,免疫鉴定法,电泳鉴定法,生物效价鉴定法,单纯指标鉴定法,细胞生物学鉴定法,DNA遗传标记鉴定法,mRNA差异显示鉴定法,聚合酶链式反应,(Polymerase Chain Reaction),简称PCR,是一种分子生物学技,术,用于放大特定的DNA片段,可看作生物体外的特殊DNA复制。,少量DNA样品快速扩,增amp|fy技术,(invented by Kary,Mullis won a Nobel Prize,in Chemistry in 1993),原理,Sequencing,1Oeyelus of& HIepw:,I: denaturation,I inur oc,ImrmmtmTmTmmn m ITITTTImTIFITT ITTITITTITITTTTTTTTTTITT,iuumLLLILunLAL,stop: annealing,aLaiN,I seconds 50c,4、f,d minnie nf i,y山uuuu山y,PCR要件,模板 Template,引物 Primers,Small pieces of DNa,Made up of 15 to 30 bases,Binds the dNa at a specific region,forward and reverse正向和逆向,酶 Enzyme,Tag DNA polymerase,PCR核酸 Nucleotides,Substrate, four deoxyribonucleotides(A, C,G,T),dNTP,Dist H2O 117 ul,10X buffer 15 u,dNTP,Primer 1,3 ul,Primer 2 3 u,Tag polymerase 0.75 ul,200ul tube,Pipette上下混合,取出15u,不加模板的对照组,剩下的加入3 ul Template DNA,用 pipette上下混合,分装入tube15u/支,标上1.5,C,54C56,62,55C,Run Pcr,不加引物但有模板的对照,加入1u模板DNA,15 ul,template,UW,1 ul,TC,3 ul template(T),Pipette mix,15ul/tube,1/3/3/45,PC,