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Position: Home > Articles > Optimization on RAPD reaction system of Rhododendron schlippenbachin Maxim Agricultural Science Journal of Yanbian University 2009,31 (1) 21-25

大字杜鹃RAPD反应体系的优化

作  者:
姬文秀;金东淳;李虎林;鲁艺
单  位:
延边大学农学院农学系
关键词:
大字杜鹃;RAPD;反应体系;优化
摘  要:
以大字杜鹃为材料,以改进的CTAB法提取大字杜鹃叶片总DNA,分别就模板DNA浓度,引物浓度,DNTP浓度,Taq DNA聚合酶量及镁离子浓度对大字杜鹃RAPD反应结果的影响进行研究.通过对各因子的组合比较,建立了大字杜鹃RAPD反应的优化体系,即总反应体积为25μL,其中包括10×Taq酶配套缓冲液2.5μL,模板DNA浓度50 mg/L,引物浓度0.5μmol/L,dNTP浓度0.15 mmol/L,Taq DNA聚合酶1 U,Mg2+浓度2.0 mmol/L,其余用重蒸馏水补足.扩增程序为94℃预变性5 min,94℃变性1 min,38℃退火40 s,72℃延伸2min,共41个循环,72℃延伸7 min.
译  名:
Optimization on RAPD reaction system of Rhododendron schlippenbachin Maxim
作  者:
JI Wen-xiu, JIN Dong-chun, LI Hu-lin*, LU Yi (Agricultural Department,Agricultural College of Yanbian University,Longjing Jiln 133400,China)
关键词:
Rhododendron schlippenbachin Maxim.;RAPD;reaction systerm;optimization
摘  要:
The genomic DNA was extracted from Rhododendron schlippenbachin Maxim.by means of advanced CTAB method.Studied the influences of the concentration of template DNA,primer,dNTP,TaqDNA polymerase and Mg 2+ on RAPD analysis procedure.Astablished the optimization system of RAPD on the base of the comparation among various influence factor groups:total volume 25 μL,comprising of 10×Taq buffer(2.5 μL),template DNA concentration 50 mg/L,primer concentration 0.5 μmol/L,dNTP 0.15 mmol/L,TaqDNA polymerase 1 U,Mg 2+ concentration 2.0 mmol/L and the supplementary ddH2O.The extended program was: 94 ℃ predenaturation 5 min,94 ℃ denaturation 1 min,38℃ annealing 40 s,72 ℃ extension 2 min,41 cycles in all,72 ℃ extension 7 min finally.

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