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首页> 外文期刊>Journal of Forestry Research >DNA extraction of birch leaves by improved CTAB method and optimization of its ISSR system
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DNA extraction of birch leaves by improved CTAB method and optimization of its ISSR system

机译：改进CTAB法提取白桦叶DNA及其ISSR体系优化。

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摘要

The basic method of DNA extraction (CTAB) was improved as the multi-times STE-CTAB extraction method and used to extract the DNA of birch leaved in this experiment. Results showed that the improved method is suitable not only for genomic DNA extraction of birch but also for that of other plants. The purity of genomic DNA extracted by the multi-times STE-CTAB extraction method is higher than that by one time STE-CTAB method, and it does not need the process of RNase. The factors of influencing ISSR system were explored based on the genomic DNA of birch extracted by the two methods. The optimal conditions for ISSR system were determined as follows: Mg~(2+) concentration is 1.5-3.0 mmol·L~(-1), dNTP concentration 0.10-0.25 mmol·L~(-1), the quantity of Taq polymerase 0.5-2.0 U, template DNA 30-100 ng, and the concentration of primer is 0.2-0.4 μmmol·L~(-1), and the reaction program was as: initial denaturation for 5 min at 94℃, 30 cycles of denaturation for 30 s at 94℃, annealing for 30 s at 51℃, extension for 30 s at 72℃, and a final 7 min extension at 72 ℃.
机译：DNA提取的基本方法（CTAB）是多次STE-CTAB提取方法的改进，并用于提取本实验中留下的桦木DNA。结果表明，改进后的方法不仅适用于桦树基因组DNA的提取，还适用于其他植物的提取。多次STE-CTAB法提取的基因组DNA的纯度高于一次STE-CTAB法提取的基因组DNA的纯度，不需要RNase的处理。基于两种方法提取的桦树基因组DNA，探讨了影响ISSR系统的因素。确定ISSR体系的最佳条件如下：Mg〜（2+）浓度为1.5-3.0mmol·L〜（-1），dNTP浓度为0.10-0.25mmol·L〜（-1），Taq聚合酶的量0.5-2.0 U，模板DNA 30-100 ng，引物浓度为0.2-0.4μmmol·L〜（-1），反应程序为：94℃初始变性5 min，变性30次在94℃下放置30 s，在51℃下退火30 s，在72℃下延伸30 s，最后在72℃下延伸7 min。

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