| 灰色家鸽ISSR反应体系的建立与优化 |
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| 引用本文： | 杨洪雁,刘诚刚,白秀娟.灰色家鸽ISSR反应体系的建立与优化[J].中国畜牧兽医,2012,39(5):44-47. |
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| 作者姓名： | 杨洪雁 刘诚刚 白秀娟 |
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| 作者单位： | 东北农业大学动物科学技术学院,黑龙江哈尔滨,150030 |
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| 摘 要： | 本研究旨在通过正交优化得出灰色家鸽的ISSR-PCR反应的最佳体系。以灰色家鸽为试验材料,使用引物ISSR807,采用正交优化方法对影响PCR反应的Mg2+、dNTPs、引物、Taq DNA聚合酶、模板进行了体系优化,同时对引物的最佳退火温度进行了选择,建立了适合灰色家鸽的ISSR-PCR分析的最佳体系。结果表明,灰色家鸽ISSR-PCR反应的最优体系为:25 μL总体积中包括10×PCR Buffer 2.5 μL, Mg2+ 2.6 mmol/L, dNTPs 0.2 mmol/L, 引物0.4 μmol/L,Taq DNA聚合酶0.5 U,DNA 模板1.2 ng/μL,最佳退火温度为56.2 ℃。通过对ISSR-PCR反应体系的优化,能为利用该技术进行灰色家鸽遗传多样性评价、不同种源鉴定及亲缘关系分析奠定基础。
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| 关 键 词： | 灰色家鸽 正交设计 体系优化 ISSR-PCR体系 |
| 收稿时间： | 2011-11-08 |
Establishment and Optimization of ISSR-PCR Reaction System for Columba domestica |
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| YANG Hong-yan
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LIU Cheng-gang
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BAI Xiu-juan.Establishment and Optimization of ISSR-PCR Reaction System for Columba domestica[J].China Animal Husbandry & Veterinary Medicine,2012,39(5):44-47. |
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| Authors: | YANG Hong-yan LIU Cheng-gang BAI Xiu-juan |
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| Institution: | College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China |
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| Abstract: | This experiment was conducted to obtain the suitable ISSR-PCR reaction system of Columba domestica by orthogonal design.Columba domestica was chosen to study the affecting factors of ISSR-PCR amplification.Orthogonal design of five factors(Mg2+,dNTPs,primer,Taq DNA polymerase,template) at four levels was used to establish the suitable ISSR-PCR system by primer ISSR807 in Columba domestica,and the suitable anneal temperature was yielded from gradient PCR on temperature.The results showed that the optimized ISSR-PCR system(25 μL reaction volume) in Columba domestica was proved to be: 10×PCR Buffer 2.5 μL,2.6 mmol/L Mg2+,0.2 mmol/L dNTPs,0.4 μmol/L primer,0.5 U Taq DNA polymerase,DNA template 1.2 ng/μL;and suitable anneal temperature was 56.2 ℃.According to the optimum ISSR-PCR reaction system,the results lay a foundation for the analyses of genetic diversity,germplasm resources and genetic relationship of Columba domestica. |
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| Keywords: | Columba domestica orthogonal design optimization ISSR-PCR system |
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