| 刘姗1,2 高玉莲1 孙蓉1 唐自钟1 高静雷1 陈惠1.金龙胆草SRAP 银染反应体系的建立与优化[J].广东农业科学,2014,41(1):121-126 |
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金龙胆草SRAP 银染反应体系的建立与优化 |
| Establishment and optimization of SRAP silver-stainedreaction of Conyza blinii L佴vl. |
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| DOI： |
| 中文关键词: 金龙胆草 SRAP PCR 体系优化 |
| 英文关键词: Conyza blinii Lvl. SRAP PCR system optimization |
| 基金项目:四川省科技创新苗子工程项目(2012ZZ046)攀枝
花学院校级科研项目(2012YB030) |
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| 中文摘要: |
| 建立适宜金龙胆草DNA 的SRAP-PCR 扩增体系为金龙胆草分子标记打下基础,针对常规银染方法从
是否固定银染液组成银染时间显影液组成等方面优化了金龙胆草SRAP 扩增产物的检测方法同时探索了金龙
胆草SRAP-PCR 反应程序,并对金龙胆草SRAP-PCR 反应体系的各影响因子进行优化结果建立了一个银染步骤
快速高效的银染检查方法筛选的退火温度为51.3,最佳SRAP-PCR 反应体系(15 L) 为院DNA 40 ng,引物浓度
0.7 mol/L,dNTPs 0.25 mmol/L,Mg2+1.9 mmol/L,Taq 聚合酶0.6 U,10buffer 2 L,双蒸水补足该程序和体系能很好
地满足金龙胆草基因组SRAP 扩增的要求SRAP 标记应用于金龙胆草遗传研究是可行的。 |
| 英文摘要: |
| To establish appropriate DNA SRAP-PCR amplification system of Conyza blinii Lvl, and lay the foundation
for the molecular markers of C. blinii Lvl. four factors of the silver-stained were optimized, including whether fixed, dye
recipe, silver staining time and composition of developer. This research also explored the SRAP-PCR reaction process and
optimized the amplification system of C. blinii by various factors. The results showed that the annealing temperature of SRAPPCR
reaction program was 51.3, and the optimum SRAP-PCR system (15 L) was as follows: DNA 40 ng, primer 0.7 mol/
L, dNTPs 0.25 mmol/L, Mg2+ 1.9 mmol/L, Taq polymerase 0.6 U, 10buffer 2 L. The program and system were suitable for
silver-stained analysis and were applicable in C. blinii L佴vl. with SRAP marker. |
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