【摘要】 以采自湖北省神农架地区的14株香果树Emmenopterys henryi为材料,提取其叶片基因组DNA,并以其基因组DNA为模板进行随机扩增多态性DNA(RAPD)反应条件的优化。RAPD反应条件中的各个因子,包括模板DNA质量浓度、引物浓度、dNTP浓度、DNA聚合酶浓度和Mg2+浓度。结果表明,香果树基因组DNA在以下条件有较好的扩增效果:25μL体系中,Taq酶1.33×10-3kat.L-1;随机引物0.5μmol.L-1;Mg2+2.6 mmol.L-1;dNTP 220μmol.L-1;DNA模板4.40 mg.L-1。聚合酶链式反应(PCR)程序为:94℃预变性5 min,94℃变性1 min,43℃退火1 min,72℃延伸2 min,经过40个循环,最后72℃延伸8 min。此体系和反应程序可获得比较稳定的扩增结果。图6表1参8更多还原

【Abstract】 Genome DNA from leaves of the Emmenopterys henryi tree(Henry emmenopterys)in the Shennongjia area of Hubei Province,China,was extracted to optimize its random amplified polymorphic DNA(RAPD)reaction conditions,which included concentrations of DNA polymerase,primers,Mg2+,dNTP,and DNA.Results indicated that genome DNA of E.henryi had good amplification with the following conditions:25 μL solution with 1.33×10-3 kat·L-1 Taq DNA polymerase,0.5 μmol·L-1 random primer,2.6 mmol·L-1 Mg2+,220 μmol·L-1 dNTP,and 4.40 mg·L-1 genomic DNA.The polymerase chain reaction(PCR) amplification program was as follows:predenaturing at 94 ℃ for 5 min,followed by 40 cycles of denaturing at 94 ℃ for 1 min,annealing at 43 ℃ for 1 min,extension at 72 ℃ for 2 min,and final extension at 72 ℃ for 8 min.Such a system and reaction programs could achieve comparatively stable amplification.[Ch,6 fig.1 tab.8 ref.]更多还原