Home Journals Chinese Agricultural Science Bulletin
Chinese Agricultural Science Bulletin

Abbreviation (ISO4): Chin Agric Sci Bull      Editor in chief: Yulong YIN

About  /  Aim & scope  /  Editorial board  /  Indexed  /  Contact  / 
目次

Establishment and Optimization of RAPD Reaction System in Tobacco

Expand

Online published: 2005-05-04

Abstract

The factors influencing RAPD Analysis, including the concentration of DNA template, Mg2+, dNTP, primers, Taq polymerase, Thermal cycles and annealing temperature in tobacco were studied. An optimal PCR system for RAPD in tobacco was found: in 25μl reaction solution, contained 40ng DNA template, 0.4μM random primers, 2.5mM Mg2+, 0.2mM dNTP, 1U Taq polymerase. The amplification program was devised: 94℃ for 5min;denaturing at 94℃ for 1min; primer annealing at 38℃ for 1min, extension at 72℃ for 1.5min, 39 cycles; at last extension at 72℃ for 5min.

Cite this article

Yang Youcai, Zhou Qingmin, Yin Hanqi . Establishment and Optimization of RAPD Reaction System in Tobacco[J]. Chinese Agricultural Science Bulletin, 2005 , 21(5) : 97 -97 . DOI: 10.11924/j.issn.1000-6850.050597

References

1Williams J G, Kubelik A R, Livak K J, et al. DNA polymorphism amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res,1990,18:6531~6535
2Welsh J M, Acclelland. Fingerprinting genomes using PCR with arbitrary primers. Mucl. Acids. Res, 1990,18:7213~7218
3李云海,钱前,曾大力,等.我国主要杂交稻亲本的RAPD鉴定及遗传关系研.作物学报,2000,26(2):171~176
4王晓武,方智远,孙培田,等.一个与甘蓝显性雄性不育基因连锁的RAPD标记.园艺学报,1998,25(2):197~198
5李海真,许通,武峻新,等.南瓜属三个种的亲缘关系与品种的分子鉴定研究.农业生物技术学报,2000,8(2):161~164
6Levi A, Thomas C E, Zhang X, et al. A genetic linkage map for watermelon based on randomly amplified polymorphic DNA markers.J. Amer. Soc. Hort. Sci,2001,126(6):730~737
7傅荣昭,孙勇如,贾士荣.植物遗传转化技术手册.北京:中国科学技术出版社,1994.131~136
8萨姆布鲁克J,弗里奇E F,曼尼阿蒂斯T.分子克隆实验指南.第二版.金冬雁,黎孟枫泽.北京:科学出版社,1992.954~960
9张菊平,巩振辉,张大过,等.苦瓜RAPD分析体系的优化研究.河南农业大学学报,2003,37(1):49~53
10 袁庆华,桂枝,张文淑.苜蓿基因组DNA提取和RAPD反应条件优选.草地学报,2001,9(2):99~105
11 Lowe A J. Standardization of molecular genetic techniques for the characterization of germplasm collections:the cases of random amplified polymorphic DNA(RAPD).Plant Gentic Resources Newsletter,1996,107:50~54
12 钟军.芸芥抗油菜菌核病分子基础的研究.[学位论文].长沙:湖南农业大学,2003.57~58
13 周章乐,王振英,彭永康.影响RAPD实验结果的稳定性和可靠性因素研究.华北农学报,2003,18(3):81~83
14 Hadrys H, Balik M, Schierwater B. Application of random amplified polymorphic DNA(RAPP) in molecular ecology. Moleculer Ecology, 1992,1:55~63
15 Bielawski J P. Noack K. Pumo D E. Reproducible Amplification of RAPD markers from Verte brate DNA. Bio Techniques, 1995,18:860~865
Options
Outlines

/