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异硫氰酸胍法快速提取二球悬铃木组织总RNA的研究 被引量:19

Guanidine Thiocyanate Method of Total RNA Isolation in Platanus aceriflia
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摘要 针对二球悬铃木组织中多酚物质含量较高的特点,采用异硫氰酸胍法从二球悬铃木花序和叶片中提取到质量高、完整性好的总RNA,28S rRNA的亮度约为18S rRNA的2倍,通过RT-PCR克隆到二球悬铃木中与拟南芥Leafy基因同源的部分编码区。高质量的RNA为Northern杂交和利用同源序列法克隆二球悬铃木的相关基因提供了前提条件。 Guanidine Thiocyanate method was used to isolate total RNA from the inflorescence tissue and leaves of Platanus aceriflia successfully with regard to high quantity of phenolic compounds in its tissue. The high-quality total RNA was obtained by guanidine thiocyanate that eliminated the interferences of polyphenols and acquired the 28S rRNA,which is approximately two times as light as the 18S rRNA. The isolated RNA was of sufficient quality for cloning partial cDNA of Leafy gene homologous to Leafy in Ara- bidopsis thaliana according to RT-PCR. The method described here was the basis of Northern blotting and cloning genes from P. aceriflia according to homologous sequence.
出处 《武汉植物学研究》 CSCD 北大核心 2007年第3期266-269,共4页 Journal of Wuhan Botanical Research
基金 国家自然科学基金资助项目(30500394) 教育部新世纪人才计划(NCET-04-0733)资助项目
关键词 二球悬铃木 多酚 异硫氰酸胍法 总RNA提取 Platanus aceriflia Polyphenol Guanidinium thiocyanate Total RNA extraction
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参考文献7

  • 1Trizol Reagent.Product description TRIZOL reagent[Z].Manufacturer protocol (1995).Life Technologies,Gaithersberg,MD.
  • 2Michael W Frohlich,David S Parker.The mostly male theory of flower evolutionary origins:from genes to fossils[J].Systematic Botany,2000,25(2):155-170.
  • 3Chomczynski P,Sacchi N.Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction[J].Anal Biochem,1987,162:156-159.
  • 4Maliyakal E J.An efficient method for isolation of RNA and DNA from plants containing polyphenolics[J].Nucleic Acids Research,1992,20(9):238-241.
  • 5Xu Q,Wen X P,Tao N G,Hu Z Y,Yue H L,Deng X X.Extraction of high quality of RNA and construction of a suppression subtractive hybridization (SSH) library from chestnut rose (Rosa roxburghii Tratt)[J].Biotechnology Letters,2006,28:587-591.
  • 6李菁芳,黄劭毅,田仁鹏,张珞珍,方呈祥.一种适用于RT-PCR的杉树类植物中总RNA提取的方法[J].武汉植物学研究,2004,22(6):551-556. 被引量:17
  • 7王曼玲,朱虹琳,周明全,胡中立,宋运淳.莲藕组织总RNA的快速提取方法[J].武汉植物学研究,2005,23(5):475-477. 被引量:22

二级参考文献24

  • 1彭秀玲 袁汉英 谢毅.基因工程实验技术[M].长沙:湖南科技出版社,1998.183-188.
  • 2Sambrook J, Maniatis T, Fritsch E F. Molecular Cloning:A Laboratory Manual[M]. 2nd ed. New York: Cold Spring Harbor Laboratory Press, 1989.343-361.
  • 3Chomczynski P,Sacchi N.Single-step method of RNA isolation by acid guanidinium thiocyanate-thiocyanate-phenol-ch-loroform extraction[J].Anal Biochem,1987,162:156-159.
  • 4Bekesiova I,Nap J P,Mlynavoval K.Isolation of high quality DNA and RNA from leaves of the carnivorous plant droscra rotwndifolia[J].Plant Mol Bio Rep, 1999,17:269-277.
  • 5Chang S, Puryear L, Caivney J. A simple method for isolating RNA from pine trees[J]. Plant Mol Bio Rep,1993,11:113-119.
  • 6Woodhead M, Taylor M A, Davies H V, Brennan R M , McNicol R J. Isolation of RNA from blackcurrant (Ribes nigrum L.) fruit[J]. Mol Biotechnol, 1997, 7(1):1-4.
  • 7Maliyakal E J. An efficient method for isolation of RNA and DNA from plants containing polyphenolics[J]. Nucleic Acids Research,1992, 20(9): 238-241.
  • 8Sambrook J,Russell D W.Molecular Cloning:A Laboratory Manual[M].3rd ed.USA:Cold Spring Harbor Laboratory Press,2001.
  • 9Su X,Gibor A. A method for RNA isolation from marine macroalgae[J].Anal Biochem,1988,174:650-657.
  • 10Katterman F R H,Shttuck V I. An effective method of DNA isolation from the matureleaves of Gossypium species that contain large amount of pheolic terpenoids and tannins[J].Prep Biochem,1983,13:347-359.

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