欢迎来到天天文库
浏览记录
ID:5355461
大小:575.84 KB
页数:4页
时间:2017-12-08
《普通甲醛固定石蜡包埋组织dna提取方法的探讨》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、万方数据342《癌症》chineseJoumalofcancer,2004,23(3):342—345普通甲醛固定石蜡包埋组织DNA提取方法的探讨田子强,刘俊峰,张少为,李保庆,王福顺,张月峰AStudyOntheMethOdOfDNAExtraCtiOnfrOmUnbufferedF0rma
2、-n-FixedandParaffin-EmbeddedSampIesTianZi—Qiang,LiuJun—Feng.ZhangSha0一Wei,LiBao—Qing,WangFu—Shun,ZhangYue—Feng河北医科大学第四医院胸外科,河北石家庄050叭lD印onm饥£矿孔or口cic5ur
3、ge佛FouTthHospt£qt《HebeiMed记nlUn西e瑚i£竹S危彬。如M。昭,胁6ei,0500,J,P.尼C^inⅡ通讯作者:刘俊峰Correspondenceto:LiuJun-FengTel:86—3ll一782667686—013931152296E—mail:tizql2@heinfo.nettizql2@vip163.com收稿日期:2003.05—20修回日期:2003—09一08·技术与方法·【ABSTRACT】BACKGROUND&OBJECTlVE:UnbufferedformaIiniswide
4、yused10flxresectedsDecimensinCh
5、ina.TheDNAinunbu什eredformalin—fixedandparaffin—embeddedtissuesisusua
6、IydegradedseriousIy,so¨1eextractiOnOfDNAfrOm甘1esesamplesisdifficuIt.ThissludywasconductedtOseekanOptimalmethOdtOextractDNAfrOmthesesamDles.METHODS:FifleenblocksofesOphageaIcarcinOmaresectedinFOurthH0spilal0fHebeiMedicaIUniversilVin
7、2000wereselected.ThecelIswerelyzedbyprOteinaseKdigestiOnOrheatingunderdi仟erentpHvaIues,thenDNAwase×tractedbyphenOl:chlOrOform.Afterthat.fourparameters(deparaffinedbyxyIeneorhistolene;digestedfor48hor72hat37℃or56’C:extractedbysaltjng—outorphenoI:chJoroform)wereOptimizedaccordingtotheprIncipIeOfcrossd
8、esign.AtIast,thequaIityofobtainedDNAwasanaIyzedw.theIectrophOresisandPCRamplificatiOn.RESULTS:ThequaIityandquantityOfDNAObtainedbyproteinaseKdigestion(theaVerageyieIdis17.88ug)werebetterthanthatofheatingunderdifferentpH(7—12)(P<0.05).ThequaIItyandquantjtyofDNAdigestedat56℃werebetterthanthatat37℃,and
9、simi
10、arIy.digestionfor72hOurswasbetterthanthatfOr48hours.ThemethOdsOfdeparaffinandextracliOnhadn0ObviOusinfluenceonthequaIityandquantityofDNA.CONCLUSION:BymeansOfNaCIsa⋯ng—outafterproteinaseKdlgestion.mOrereIiablequaIjtyOfDNAcanbeobtainedfromunbu仟eredfOrmalin一佰xedandparaffln.embeddedsamples.Furtherm
11、Ore,djgestiOnforlhreedaysat56℃ismoreIIkelytOobtainDNAwithhighqualityandquantity.KEYWORDS:EsOphageaIneOplasms:Tumortissues:Paraffin—embedded:FormaJIn—fixed:DNAextraction:PoIymerasechajnreaction(PC只)【摘要
此文档下载收益归作者所有