HLA分型技术;Variation1%atasinglegeneticlocusinapopulationofindividuals
Eachpolymorphicvariantiscalledanallele
Inthehumanpopulation,over1,695MHCalleleshavebeenidentified;Polymorphicresiduesarelocatedintheantigen-bindinggroove
—Variationinaminoacidsequenceschangestheshapeofthegroove;HLA分型概述;PCR-SSP检测HLA-B27;EveryHLAallelehasitsownuniquesequences;Specificsequenceprimer(SSP)
isdesignedtobindtotheallelicsequencescomplementarily;SequencespecificprimerisusedtotypeHLAallelewithPCR;;实验步骤;三、实验材料和方法
1、血样:0.2mlEDTA抗凝血
2、红细胞裂解液
3、白细胞裂解液
4、PCR混合液
PCRbuffer
HLA-B27SSP
Taq
dNTP
internalpositivereferenceprimers
;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;SpecificbandscanbeseenwithHLA-B27(+)samples;谢谢聆听!