Safer DNA extraction from plant tissues using sucrose buffer and glass fiber filter
Safer DNA extraction from plant tissues using sucrose buffer and glass fiber filter
复制标题
使用蔗糖缓冲液和玻璃纤维过滤器从植物组织中更安全地提取 DNA
DOI：
10.1007/s10265-012-0502-x
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发表时间：
2012
影响因子：
2.8
通讯作者：
Koh-ichi Takakura & Takayuki Nishio
中科院分区：
文献类型：
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作者：
Koh-ichi Takakura & Takayuki Nishio
对于一些植物物种，DNA提取和下游实验受到多种化学物质如多糖和多酚的抑制。本文提出了一种不含有机溶剂（乙醇除外）的提取方法。该方法由STE缓冲液（0.25 M蔗糖，0.03 M Tris, 0.05 M EDTA）的初始洗涤步骤组成，然后使用一块玻璃纤维过滤器提取DNA。该方法的优点是安全、成本低。这种方法得到的DNA溶液的纯度不一定比STE/CTAB法高，但对于PCR实验来说是足够的。在过去的研究中，这些观点被证实是用两种物种，日本草蒲公英和普通蒲公英的DNA难以通过PCR扩增。
For some plant species, DNA extraction and downstream experiments are inhibited by various chemicals such as polysaccharides and polyphenols. This short communication proposed an organic-solvent free (except for ethanol) extraction method. This method consists of an initial washing step with STE buffer (0.25 M sucrose, 0.03 M Tris, 0.05 M EDTA), followed by DNA extraction using a piece of glass fiber filter. The advantages of this method are its safety and low cost. The purity of the DNA solution obtained using this method is not necessarily as high as that obtained using the STE/CTAB method, but it is sufficient for PCR experiments. These points were demonstrated empirically with two species, Japanese speedwell and common dandelion, for which DNA has proven difficult to amplify via PCR in past studies.