河南农业大学学报2011，Vol.45Issue(5)：523-529,7.
蜡梅AFLP反应体系优化及引物筛选
Optimization of Chimonanthus praecox AFLP reaction system and its primer selection
摘要
Abstract
The chimonanthus praecox AFLP system was established and its primer was selected for analysis, by optimizing the main factors affecting the AFLP system. The results indicated that the AFLP optimum conditions for restriction enzyme digestion of the Chimonanthus praecox were as follows: 600 ng DNA template, 3 U Pst I and Mse I respectively in 20 Μl reaction volume , at 37 ℃ for 2 h; The ligation system (20 Μl) with 15 Μl the digestion product, 3 U T4 ligation enzyme , 0. 25 (μmol · L-1 Pst I adapter, 2.5 μmol · L-1 Mse I adapter, 1 Μl 10 × T4 Buffer, at 22 ℃ for 12 h was the optimum condition; The preamplification reaction (20 Μx) with 5 Μl the ligation fragment which was diluted 10 times, 2.0 mmol · L-1 Mg2+ , 2 U Taq enzyme, 300 μmol · L-1 Dntp, 0.5 μmol · L-1 Pst I and Mse I primer ( P + AGA/M + ATC ) was the optimum condition; The selective amplification reaction (20 Μl) with 5 Μl preamplification product which was diluted 20 times, 2.0 mmol · L-1 Mg2+ , 2 U Taq enzyme, 300 (μmol · L-1 Dntp, 0. 5 μmol · L-1 Pst I and Mse I primer (P + AGA/M + ATC ). Finally, 96 pairs of primers were screened out for the AFLP analysis of Chimonanthus praecox plants.关键词
蜡梅/AFLP/体系优化/引物筛选Key words
Chimonanthus praecox/AFLP/reaction system/primer selection分类
农业科技引用本文复制引用
赵明晓,冯志敏,胡湛,赵振利,范国强..蜡梅AFLP反应体系优化及引物筛选[J].河南农业大学学报,2011,45(5):523-529,7.基金项目
林业公益性行业科研专项(200904009) （200904009）
