【摘要】 为进一步开展石蒜属Lycoris植物种质资源遗传多样性研究,利用正交试验设计的方法,从Taq酶、Mg2+、模板DNA、dNTP和引物等5因素4水平,对石蒜属植物ISSR(intersimple sequence repeats)反应体系进行优化,确立了石蒜属植物ISSR的最佳反应体系:在20μL反应体系中,含1×Buffer,模板DNA 50 ng,2.5 mmol.L-1氯化镁(MgCl2),0.15 mmol.L-1dNTP,25.05 nkatTaq聚合酶,0.5μmol.L-1引物。进一步进行梯度退火试验,确定最适宜退火温度为57℃。图7表2参23更多还原

【Abstract】 To study the genetic diversity of Lycoris Herb.(Amaryllidaceae),an orthogonal design was used to optimize an inter simple sequence repeats(ISSR) amplification system on Lycoris using five factors(Mg2+,deoxyribonucleotide triphosphate(dNTP),primers,DNA template,and Taq polymerase) at four levels of concentrations respectively.A suitable ISSR reaction system was constructed with the 20 μL reaction system containing 1 × polymerase chain reaction(PCR) buffer,2.5 mmol·L-1 MgCl2,150 μmol·L-1 dNTP,0.5 μmol·L-1 primer,50 ng DNA template,and 25.05 nkat Taq DNA polymerase.According to the gradient PCR,the optimal annealing temperature for the ISSR reaction was 57 °C.[Ch,7 fig.2 tab.23 ref.]更多还原