莲雾ISSR反应体系的优化与应用

以黑珍珠莲雾为试材,对ISSR反应体系中的各个主要影响因子进行了优化筛选,结果表明,25μL ISSR反 应体系各组分的最适浓度分别为:1×Buffer(不含Mg2+)、2.0 mmol/L Mg2+、0.2 mmol/L dNTPs、Taq DNA聚合酶1.5 U、引 物0.2 μmol/L、模板1.0 ng/μL。此外,加入1.6％的甲酰胺有利于减轻背景噪音的干扰。并利用该优化体系对12个连 雾类型和2个近缘种进行了ISSR扩增,证实了该体系稳定可靠。

Optimization of the inter-simple sequence repeat reaction system and its preliminary application in wax apple(Syzygium samarangens)

The optimization of main influential factors of ISSR reaction system in wax apple(Syzygium samarangens), black pearl cultivar was studied. The results showed that the optimum concentrations of components such as Buffer, Mg2+,dNTPs, Taq DNA polymerase, primer, template DNA in 25 mL reaction system were 1×Buffer( without Mg2+), 2.0 mmol/L,0.2 mmol/ L, 1.5 U, 0.2 μmol/L, 1.0 ng/mL, respectively. Furthermore, the addition of 1.6% formamide was propitious to reduce the disturbance. Amplifications of 12 varieties of wax apple and two closely related species were achieved using this optimum system.