普通甲醛固定石蜡包埋组织dna提取方法的探讨

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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