四川农业大学学报 ›› 2004, Vol. 22 ›› Issue (01): 30-33.doi: 10.16036/j.issn.1000-2650.2004.01.009

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金花梨基因组DNA提取及其RAPD分析

刘遵春, 廖明安, 任雅君   

  1. 四川农业大学 林学园艺学院, 四川 雅安 625014
  • 收稿日期:2003-12-18 出版日期:2004-03-31 发布日期:2017-03-04
  • 基金资助:
    四川省学术和技术带头人培养资金资助项目

Genome DNA Extraction and RAPD Analysis of Jinhua Pear

LIU Zun-chun, LIAO Ming-an, REN Ya-jun   

  1. College of Forestry and Horticulture, Sichuan Agricultural University, Yaan 625014, Sichuan, China
  • Received:2003-12-18 Online:2004-03-31 Published:2017-03-04

摘要: 以金花梨嫩芽为试材,通过在磨样时添加不同量的抗氧化剂Vc,对CTAB法作了一定的改进,比较了不同处理的提取效果,再分别以成熟叶片、新梢和嫩芽为材料,采用改进的CTAB法提取基因组DNA,进一步比较了不同材料的提取效果,并以金花梨及其18个变异单系的基因组DNA为模板,40个随机引物进行RAPD分析。结果表明:以嫩芽为试材,在磨样时每克鲜样添加0.10g的Vc,能够提取到高质量的基因组DNA;采用改进的CTAB法,用成熟叶片、新梢和嫩芽均能提取到可直接用于RAPD分析的基因组DNA;大部分引物可以在不同模板上扩增出条带,但仅有5个引物可以同时在金花梨及其18个变异单系基因组DNA上扩增出条带;用RAPD结果对金花梨及其18个变异单系进行聚类分析。

关键词: 金花梨, DNA提取, RAPD分析

Abstract: The CTAB method was definitely improved by different doses of Vc appended when the sample of tender bud of Jinhua pear was triturated. And then the isolation effect of genomic DNA from mature leaf, shoot and tender bud of Jinhua pear was studied respectively by improved CTAB method. After comparing the quantity and quality of genomic DNA extracted from different materials, the high-quality genome DNA of Jinhua pear and its 18 varieties was used as templates and amplified with 40 random primers by using RAPD. The result shows that with the dose of 0.10 g/g of Vc added when the sample is triturated, the high-quality of genomic DNA can be extracted from tender bud, and the genomic DNA can be extracted from mature leaf, shoot and tender bud all can be used for RAPD analysis. Fragments are amplified on different templates with most of 40 random primers. Only five primers get better result on all templates. According to the results of RAPD, the genetic relationship among Jinhua pear and its 18 varieties is examined for classification of Jinhua pear.

Key words: Jinhua pear, DNA extraction, RAPD analysis

中图分类号: 

  • S661.2