| 蜜蜂遗传多样性研究的RAPD-PCR反应体系的正交优化 |
| |
| 引用本文： | 刘楠楠,薛运波,王志,李志勇.蜜蜂遗传多样性研究的RAPD-PCR反应体系的正交优化[J].吉林畜牧兽医,2011(9):4-7. |
| |
| 作者姓名： | 刘楠楠 薛运波 王志 李志勇 |
| |
| 作者单位： | 吉林省养蜂科学研究所; |
| |
| 基金项目： | 国家蜂产业技术体系建设专项经费项目资助(编号:CARS-45-KXJZ); 吉林省科技发展计划项目(编号:20080559) |
| |
| 摘 要： | 利用正交设计L16（45）对蜜蜂基因组DNARAPD-PCR反应体系的5个因素（Tag酶,引物,Mg2＋,dNTP,模板）在4个水平上进行优化试验,筛选出各个反应因素的最佳水平,建立蜜蜂模板DNARAPD-PCR反应的最佳体系（25L）：10×PCRbuffer3.0L,Tag酶0.5U,引物浓度0.4mol/L,Mg2＋浓度3.0mmol/L,dNTP浓度0.5mmol/L,模板40ng。对蜜蜂DNARAPD-PCR最佳反应体系的退火温度进行了梯度试验,最佳退火温度为54℃。
|
| 关 键 词： | 蜜蜂 RAPD-PCR反应体系 正交设计 |
Optimization of RAPD-PCR Reaction System for the of Genetic Diversity Study of Honeybee |
| |
| LIU Nan-Nan,XUE Yun-Bo,WANG Zhi,LI Zhi-Yong.Optimization of RAPD-PCR Reaction System for the of Genetic Diversity Study of Honeybee[J].Jilin Animal Science and Veterinary Medicine,2011(9):4-7. |
| |
| Authors: | LIU Nan-Nan XUE Yun-Bo WANG Zhi LI Zhi-Yong |
| |
| Institution: | LIU Nan-Nan,XUE Yun-Bo,WANG Zhi,LI Zhi-Yong(Apiculture Science Institute of Jilin Province,Jilin 132108) |
| |
| Abstract: | Orthogonal design was used to optimize RAPD-PCR amplification system for Apis mellifera in terms of 5 factors（Taq DNA polymerase,Mg2＋,primer,dNTP and DNA template）from 4 levels.An optimal RAPD-PCR system for Apis mellifera was obtained as 10 × PCR buffer3.0 l,0.5U Taq DNA polymerase,0.4 mol/L primer,3.0mmol/L Mg2＋,0.5mmol/L dNTP,and 40ng DNA template for 25 l reaction system.The optimal annealing temperature for RAPD-PCR reaction system was determined as 54℃ by gradient PCR. |
| |
| Keywords: | honeybee RAPD-PCR reaction system orthogonal design |
| 本文献已被 CNKI 维普 等数据库收录！ |
|
