Acta Agrestia Sinica

›› 2002, Vol. 10 ›› Issue (1): 18-23.DOI: 10.11733/j.issn.1007-0435.2002.01.003

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Optimalization of RAPD Reaction System for Alfalfa

SUN Jie1, YANG Qing-chuan2, HAN Guo-dong 3   

  1. 1. Agricultural Polytechnic college, Chifeng 024031 China;
    2. Institute of Animal Science, CAAS, Beijing 100094 China;
    3. Ecological Environment College of Inner Monglia Agricultural University, Huhhot 010018, China
  • Received:2001-07-05 Revised:2001-10-11 Online:2002-02-15 Published:2002-02-15

紫花苜蓿RAPD反应条件优化

孙杰1, 杨青川2, 韩国栋3   

  1. 1. 内蒙古农业大学生态环境学院, 呼和浩特, 010018;
    2. 中国农业科学院畜牧研究所, 北京, 100094;
    3. 内蒙古农业大学生态环境学院, 呼和浩特, 010018
  • 作者简介:孙杰(1962- ),女,内蒙古自治区赤峰农牧学校讲师,主讲饲料生产学.现为内蒙古农业大学在职硕士生,在中国农业科学院畜牧所参加紫花苜蓿耐盐遗传育种研究.
  • 基金资助:
    国家自然科学基金项目(39870560)资助.

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Abstract: Three species of alfalfa are used as the materials of extracting DNA in this experiment, they are the Zhongmu number one?Wei nan?Atlantic ocean.The experiment step to the concentration of MgCl2,dNTP,random primers,template DNA, TaqDNA polymerase and buffer on RAPD are conduced? Then a stable reaction system of RAPD is set up.In the optimalization of RAPD system with totale volume of 25 μL including MgCl2 at 2.7 mmol/L, dNTP at 150.0 μmol/L, random primer at 0.192 μmol/L,TaqDNA polymerase at 2 u/25 uL,buffer at 50.0 mmol/L and 120.0ng template DNA.The effect of amplifing with the reaction system is the most optimal.

Key words: Alfalfa, RAPD, Amplifed reaction, Optimalization

摘要： 以"中苜一号"、"大西洋"、"渭南"3个苜蓿品种为材料,对RAPD反应条件中的MgCl2浓度、dNTP浓度、随机引物浓度、模板DNA用量、TaqDNA聚合酶用量以及扩增缓冲液浓度分别进行梯度试验.结果表明,该反应体系的体积为25μL,MgCl2浓度为2.7mmol/L,dNTP浓度为150μmol/L,随机引物浓度为0.192μmol/L,TaqDNA聚合酶用量为每一反应体系2U,扩增缓冲液浓度(以KCl浓度为指标)为50mmol/L,模板DNA用量为120ng.通过试验建立了一套稳定的RAPD反应体系.该反应体系扩增效果好.

关键词: 苜蓿, RAPD, 扩增反应, 优化

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