【分享】可以和TaqMan媲美的但更便宜SNP genotyping方法

如果大家在用TaqMan做SNPgenotyping，应该知道试剂有多贵。有兴趣可以看看这篇文章，一种新的SNPgenotyping方法。简单而实用。
Biotechniques.2005Dec;39(6):885-93.RelatedArticles,Links

High-throughputSNPgenotypingbysingle-tubePCRwithTm-shiftprimers.

WangJ,ChuangK,AhluwaliaM,PatelS,UmblasN,MirelD,HiguchiR,GermerS.

HumanGeneticsDepartment,RocheMolecularSystems,Alameda,CA94501,USA.jun.wang@roche.com

Despitemanyrecentadvancesinhigh-throughputsinglenucleotidepolymorphism(SNP)genotypingtechnologies,thereisstillagreatneedforinexpensiveandflexIBLemethodswithareasonablethroughput.Herewereportsubstantialmodificationsandimprovementstoanexistinghomogenousallele-specificPCR-basedSNPgenotypingmethod,makingitanattractivenewoptionforresearchersengagingincandidategenestudiesorfollowingupongenome-widescans.Inthisadvancedversionofthemeltingtemperature(Tm)-shiftSNPgenotypingmethod,weattachtwoGC-richtailsofdifferentlengthstoallele-specificPCRprimers,suchthatSNPallelesingenomicDNAsamplescanbediscriminatedbytheTmsofthePCRproducts.Wehavevalidated306SNPassaysusingthismethodandachievedasuccessrateinassaydevelopmentofgreaterthan83%underuniformPCRconditions.Wehavedevelopedastandalonesoftwareapplicationtoautomaticallyassigngenotypesdirectlyfrommeltingcurvedata.Todemonstratetheaccuracyofthismethod,wetyped592individualsfor6SNPsandshowedahighcallrate(>98%)andhighaccuracy(>99.9%).Withthismethod,6-10,000samplescanbegenotypedperdayusingasingle384-wellreal-timethermalcyclerwith2-4standard384-wellPCRinstruments.