中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 锥栗自然居群ISSR-PCR分析技术的建立 果树学报 2009,26 (1) 103-107
Position: Home > Articles > Establishment of inter -simple sequence repeat reaction system in Castanea henryi Journal of Fruit Science 2009,26 (1) 103-107

锥栗自然居群ISSR-PCR分析技术的建立

作  者：
刘国彬;龚榜初;罗正荣
单  位：
中国林业科学研究院亚热带林业研究所;华中农业大学园艺植物生物学教育部重点实验室
关键词：
锥栗;自然居群;ISSR
摘  要：
提取野生锥栗的基因组DNA作为ISSR-PCR模板,对反应体系中的模板用量、Mg2+浓度、dNTPs浓度、引物浓度和TaqDNA聚合酶用量5个主要影响因素进行梯度试验,并对扩增程序中的退火温度与循环次数进行了探讨,初步确立了适合野生锥栗的ISSR-PCR分析技术体系,即25μL反应体系中,模板DNA40或50ng、Mg2+2.25mmol.L-1、dNTPs0.2mmol.L-1、引物0.2μmol.L-1、TaqDNA聚合酶1.0U;PCR扩增程序为:94℃预变性3min;然后94℃变性45s,(Tm+2～4℃)退火45s,72℃延伸2min,共32个循环;最后72℃充分延伸5min;1.5%琼脂糖凝胶电泳分离扩增产物。
译  名：
Establishment of inter -simple sequence repeat reaction system in Castanea henryi
作  者：
LIU Guo-bin1,2,GONG Bang-chu1*,LUO Zheng-rong2 (1Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang, Zhejiang 311400 China; 2Key Laboratory for Horticul- tural Plant Biology, Ministry of Education, Huazhong Agricultural University, Hubei, Wuhan 430070 China)
关键词：
Castanea henryi; Natural population; ISSR
摘  要：
In order to investigate the genetic diversity and relationship within and among Castanea henryi, Zhejiang and Huangshan natural C. henryi were subjected to inter-simple sequence repeat (ISSR-PCR) analysis. Genomic DNA of C. hen- ryi was extracted from leaves as the template, the mainly influencing factors of ISSR were studied and the experiment param- eters were optimized. By adjusting the concentration of template DNA, Mg2+, dNTPs, primer and the contents of Taq DNA polymerase, the PCR amplification system were optimized; Then we also optimized the amplified program, including the an- nealing temperature and PCR cycles, the PCR amplification conditions were established, finally. The optimal experiment conditions were as follow: 25 μL reaction system containing 40 or 50 ng template DNA, 2.25 mmol·L-1 Mg2+, 0.2 mmol·L-1 dNTPs, 1.0 U Taq DNA polymerase, 0.2 μmol·L-1 primer; with a ABI thermal cycle, the optimal amplification program was an initial denature for 3 min at 94 ℃, followed by 32 cycles of 45 s at 94 ℃, 45 s at Tm+2~4 ℃, 2 min at 72 ℃, and a final extension for 5 min at 72 ℃.

相似文章

导出引用
计量
文章访问数: 12
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46