【摘要】 本研究主要对杨树RAPD体系进行优化,以银白杨为试材,利用L16(45)正交试验设计和2个单因素试验,研究各主要参数的适宜浓度。结果表明,杨树RAPD优化后的反应体系:20μL反应体系中,含Mg2+1.5 mmol/L、Taq酶2.0 U、引物0.2μmol/L、dNTPs 0.2 mmol/L、模板DNA 50 ng。扩增程序为:94℃预变性3 min、94℃变性40 s、38℃退火60 s、72℃延伸90 s,共38个循环;最后在72℃延伸7 min后4℃保温。通过1.5%的琼脂糖凝胶电泳检测其扩增产物,证明该体系稳定可靠,可以用于杨树的遗传学分析。更多还原

【Abstract】 The aim of this study was to optimize the RAPD reaction system in poplar. The L16(45) orthogonaltests design and two single factors tests were used to study the appropriate concentrations of the main factorswith Populus alba. The results showed that an optimized RAPD reaction system was established: the total20 μL contained Mg2 +1.5 mmol/L, Taq polymerase 2.0 U, primer 0.2 μmol/L, dNTPs 0.2 mmol/L, DNAtemplate 50 ng. Amplified procedure was: 94℃ for 3 min, 38 cycles at 94℃ for 40 s, 38℃ for 60 s, 72℃ for90 s, followed by 72℃ for 7 min and then ramped to 4℃ to hold the reaction. Amplified products were detectedby electrophoresis in 1.5% agarose gel. It was evident that the reaction system established in this study wasstable and reliable for RAPD analysis, and could be used on genetic analysis of poplar.更多还原