HLAB27检测（教学课件）.pptVIP

PCR-SSP检测HLA-B27 ;实验步骤 ;PCR扩增的基本原理;PCR扩增体系; Polymorphism in the MHC Variation 1% at a single genetic locus in a population of individuals Each polymorphic variant is called an allele In the human population, over 1,200 MHC alleles have been identified. MHC genes are the most polymorphic known;Polymorphic residues are located in the antigen-binding groove — Variation in amino acid sequences changes the shape of the groove;Schematic diagram of class I and class II MHC genes, mRNA transcripts, and protein molecules;Every HLA allele has its own unique sequences;Specific sequence primer (SSP) is designed to bind to the allelic sequences complementarily ;Sequence specific primer is used to type HLA allele with PCR;;三、实验材料和方法 1、血样：0.2 ml EDTA 抗凝血 2、红细胞裂解液 3、白细胞裂解液 4、PCR 混合液 PCR buffer HLA-B27 SSP Taq dNTP internal positive reference primers ;HLA-B27检测操作流程;二、PCR扩增 1、吸2ul模板DNA入装有PCR混合液的小管中（注意不要加到石蜡油层）； 2、将小管置于PCR仪内进行扩增。（需80min） ;三、电泳检测 吸PCR产物10ul加到2%琼脂糖凝胶孔内； 将琼脂糖凝胶板置电泳槽内电泳160V?20min后取出，观察结果。 ;四、结果观察 ;Specific bands can be seen with HLA-B27(+) samples ;HLA分型概述;9、 人的价值，在招收诱惑的一瞬间被决定。*** 10、低头要有勇气，抬头要有低气。**** 11、人总是珍惜为得到。***** 12、人乱于心，不宽余请。**** 13、生气是拿别人做错的事来惩罚自己。***** 14、抱最大的希望，作最大的努力。**** 15、一个人炫耀什么，说明他内心缺少什么。。***** 16、业余生活要有意义，不要越轨。*** 17、一个人即使已登上顶峰，也仍要自强不息。****;