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碩士 國立清華大學 材料科學工程學系 105 abstract hide

采用RDP试剂(改进自ChomczynskiandSacchi1987)提取锯缘青蟹精巢和卵巢的总RNA,经oligotex试剂盒纯化得到mRNA.利用SMART(switchingmechanismat5'endofRNAtranscript)技术,构建成精巢和卵巢的全长cDNA文库.在构建好的文库中,精巢的文库含有约1.45×106的重组子,卵巢的文库含有约为1.75×106的重组子.插入cDNA长度为500bp～3kb,说明两文库均具有良好的质量,为进一步筛选、克隆精巢与卵巢特异表达基因奠定了基础. 【英文摘要】 To clone the testis and ovary specific expression genes of Scylla serrata,we constructed two full-length cDNA libraries using the SMART cDNA library construction kit(Clontech).Total RNA were prepared from testes and ovaries using RDP,a reagent modified from Chomczynski and Sacchi(1987)for the simultaneous isolation of RNA,DNA,and protein.Oligotex(QIAGEN)was used to separate the mRNA of testes and ovaries from total RNA.The"anchor first-strand cDNA"containing asymmetricalSfiⅠrestriction enzyme sites(A&B) was...