濒危物种胡杨和灰叶胡杨总DNA提取及RAPD体系优化

以濒危物种胡杨和灰叶胡杨硅胶干燥的叶片为材料,研究其总DNA提取方法及RAPD-PCR体系优化条件。结果表明,用改良CTAB法提取的DNA适用于胡杨和灰叶胡杨的RAPD分析。优化的胡杨和灰叶胡杨RAPD-PCR体系为:20μL的反应体系中含Mg2+2.5 mmol·L-1、dNTP 0.15 mmol·L-1、引物0.2μmol·L-1、模板DNA100 ng和1.5 U Taq DNA聚合酶。用6条多态性引物对优化体系的稳定性进行检测,结果显示扩增结果稳定,多态性丰富。该优化体系为在分子水平开展胡杨和灰叶胡杨的保护生物学研究提供了技术支持和参考。

Total DNA Extraction and RAPD System Optimization of Endangered Plant Populus euphratica and P.pruinosa

An improved method based on CTAB was used to extract the total DNA from dry leaves of endangered plant Populus euphratica and P.pruinosa,and to research their reaction system of RAPD-PCR.The results showed that the total DNA extracted by the improved CTAB method could be fit for the RAPD analysis of P.euphratica and P.pruinosa.A suitable RAPD-PCR system for P.euphratica and P.pruinosa was developed as follows: 20μL PCR solution,including 2.5mmol·L-1 Mg2+,0.15mmol·L-1 dNTP,0.2μmol·L-1 primer,100ng DNA template and 1.5U Taq DNA polymerase.By testing the stability of the system with 6 polymorphic primer pairs,the results indicated that the developed system could amplify stably and keep a rich polymorphism.These results provided a reference for the further research on both P.euphratica and P.pruinosa at molecular level.