期刊文献+

赤眼蜂SSCP分子标记体系的优化 被引量:2

Optimization of SSCP Technique in Trichogramma spp.
下载PDF
导出
摘要 以松毛虫赤眼蜂为试验材料,对SSCP分子标记体系的电泳条件、温度、变性时间、上样缓冲液等诸多影响因素进行了研究。结果表明:4℃恒温,7W恒功率下,上样缓冲液中不添加甘油、用量为PCR产物体积的0.5~3.5倍,变性温度95℃或98℃,变性时间10~15min,电泳缓冲液为1×TBE,电泳16h,为SSCP分子标记体系最佳的条件组合。 Single strand conformation ploymorphism(SSCP)technique is easy to be affected by many factors such as loading buffer,denaturalization temperature and time,and electrophoresis factors.The influences of loading buffer,denaturalization temperature and time and electrophoresis factors(electrophoresis temperature,electrophoresis buffer and electrophoresis power)on SSCP technique were studied in Trichogramma spp,The results showed when volume of the loading buffer without glycerol was 0.5~3.5 times of the volume of the PCR buffer;denaturalization temperature was 95℃ or 98℃;denaturalization time was 10~15 minutes;electrophoresis buffer was 1 × TBE;the electrophoresis temperature was about 4℃;the electrophoresis power was 7W;the electrophoresis time was 16h;the bands of the single strand DNA were clear and easy to read.
出处 《沈阳农业大学学报》 CAS CSCD 北大核心 2011年第4期428-432,共5页 Journal of Shenyang Agricultural University
基金 国家自然科学基金项目(30871674 30971962) "十一五"国家科技支撑计划项目(2006BAD0806-01) 辽宁省高校重点实验室支持计划项目(2008S203) 辽宁省教育厅科研项目(L2010488) 沈阳农业大学教师科研基金项目(20091012)
关键词 赤眼蜂 SSCP 反应体系 Trichogramma spp. SSCP reaction system
  • 相关文献

参考文献14

  • 1HAYASHI K. PCR-SSCP: a simple and sensitive method for detection of mutations in the genomic DNA [J].PCR Methods and Applications, 1991 (1):34-38. H. 被引量:1
  • 2AYASHI K, YANDELL D W. How sensitive is PCR-SSCP[J].Human Mutation,1993,2:338-346. 被引量:1
  • 3ANTOLIN M F, BOSIO C F, COTTON J, et al. Intensive linkage mapping in a wasp (Bracon hebetor) and a mosquito (Aedes aegypti) with single-strand conformation polymorphism analysis of random amplified polymorphie DNA markers[J]. Genetics,1996,143:1727-1738. 被引量:1
  • 4ZHANG A, DUNN J B, CLARK M. An efficient strategy for validation of a point mutation associated with acetylcholinesterase sensitivity to azinphosmethyl in Colorado potato beetle[J].Pestic Bioehem Physiol,1999,65:25-35. 被引量:1
  • 5COLTON Y M, CHADEE D D, SEVERSON D W. Natural skip oviposition of the mosquito Aedes aegypti indicated by codominant genetic markers[J].Med Vet Entomol,2003,17:195-204. 被引量:1
  • 6FULTON R E, SALASEK M L, DUTEAU N M, et al. SSCP analysis of cDNA markers provides a dense linkage map of the Aedes aegypti genome[J].Geneties,2001,158:715-726. 被引量:1
  • 7URDANETA-MARQUEZ L, BOSIO C, HERRERA F, et al. Genetic relationships among Aedes aegypti collections in venezuela as determined by mitoehondrial DNA variation and nuclear single Nucleotide polymorphisms [J].Am J Trop Med Hyg,2008,78(3) : 479-491. 被引量:1
  • 8ROS V I D, BREERWER J A J. Spider mite (Aeari: Tetranyehidae) mitochondrial COl phylogeny reviewed: host plant relationships, phylogeography, reproductive parasites and barcoding[J].Exp Appl Acarol,2007,42:239-262. 被引量:1
  • 9余桂红,唐克轩,马鸿翔,周淼平,张旭,任丽娟,陆维忠.小麦SSCP分子标记体系的优化[J].核农学报,2007,21(4):333-338. 被引量:11
  • 10SUNNUCKS P, WILSON A C C, BEHEREGARAY L B, et al. SSCP is not so difficult: the application and utility of single-stranded conformation polymorphism in evolutionary biology and molecular eeology [J].Molecular Ecology,2000,9 (11): 1699-1710. 被引量:1

二级参考文献54

  • 1丁建松,曹毅,童建.PCR-SSCP研究进展[J].辐射防护通讯,2004,24(5):27-31. 被引量:4
  • 2杨志惠,周斌,贾静,王闯,张林.PCR-SSCP分析参数的研究[J].泸州医学院学报,2005,28(5):391-393. 被引量:10
  • 3姚海军,曹虹,郭辉玉.PCR—SSCP分析法及其研究进展[J].生物技术,1996,6(4):1-4. 被引量:16
  • 4方美英,姜志华,刘红林,黄月英,李齐贤,朱婉茹,李学斌,陈杰.应用PCR-SSCPs、PCR-RFLPs技术检测猪氟烷基因[J].养猪,1997(1):28-29. 被引量:14
  • 5KAKUTANI T, HAYANO Y, HAYASHI T, et al. Ambisense segment 4 of R ice s tripe virus possible evolutionary relationship with phoeboviruses and uukuviru ses (Bunyaviridae)[J]. J Gen Virol,1990, 72(2):465-468. 被引量:1
  • 6ORITA M, SUZUKI Y, SEKIYA T,et al. Rapid and sensitive detection o f point mutations and DNA polymorphisms using the polymerase chain reaction[J]. Genomics,1989,5:874-879. 被引量:1
  • 7ORITA M,IWAHANA H, KANAZAWA H, et al. Detection of polymorphisms of human DN A by gel electrophoresis as single-strand conformation polymorphisms[J]. Proc Natl Acad Sci USA, 1989,86:2766-2770. 被引量:1
  • 8HAYASHI K, YANDELL DW. How sensitive is PCR-SSCP[J]. Hum Mut,1993 ,2:338-346. 被引量:1
  • 9NAKAMURA H, KANEKO S, YAMAOKA Y, et al. PCR-SSCP analysis of the r ibosomal D NA ITS regions of the willow rust fungi in Japan[J]. Ann Phytopathol Soc Jpn, 1998,64:102-109. 被引量:1
  • 10Noumi T.,Mosher M.E.,Natori S.,Tutai M.,Kanazawa H.J.Biol.Chem.,1984,259:10 071~10 075. 被引量:1

共引文献56

同被引文献11

引证文献2

二级引证文献2

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部