Background:Research focused on extreme environments is often associated with difficulties in obtaining fresh plant material. Herbaria may provide great support as they house large collections of specimens from different parts of the world. Accordingly, there is also a growing interest in methods using herbarium specimens in molecular studies. Much of the literature on herbarium DNA is aimed to improve extraction and PCR amplification and is focused mostly on vascular plants. Here, I provide a brief study of DNA extraction efficiency from moss herbarium specimens, emphasizing the importance of herbaria as an invaluable source of material from hard-to-access geographical areas, such as the Antarctic region. Methods:The presented study is based on herbarium collections of 25 moss species collected in the austral polar regions between 1979 and 2013. The majority of samples were obtained using the DNeasy Plant Mini Kit (Qiagen, Hilden, Germany). The remaining, smaller part was extracted using an adapted CTAB-based approach. The performance of DNA extraction methods in terms of PCR amplification success was measured by testing several DNA fragments of various size. Furthermore, in order to estimate of DNA fragmentation level, an automated on-chip electrophoresis system was used. Results:Results reveal that DNA purity and the length of the target genetic region are the fundamental agents which drive the successful PCR reaction. Conversely, the DNA yield and specimen age seem to be less relevant. With this study, I present also an optimized CTAB-based approach which may effectively suppress inhibitors in the herbarium DNA. This method can be considered a cheaper alternative to column-based technology, particularly useful for dealing with a large number of samples. Results of this study confirmed previous reports and contribute to filling the existing gap in molecular analyses which involve the use of herbarium collections of mosses.

译文

背景:针对极端环境的研究通常与获取新鲜植物材料有关。草本植物可能会提供很大的支持,因为它们会收集来自世界各地的大量标本。因此,在分子研究中使用植物标本室标本的方法也越来越引起人们的兴趣。关于植物标本室DNA的许多文献旨在改善提取和PCR扩增,并且主要集中在维管植物上。在这里,我对从苔藓植物标本室标本中提取DNA的效率进行了简要研究,强调了草as作为难以接近的地理区域(如南极地区)的宝贵材料来源的重要性。
方法:本研究基于1979年至2013年在南极地区收集的25种苔藓植物标本室的收集。大部分样品是使用DNeasy Plant Mini Kit(Qiagen,Hilden,德国)获得的。使用基于CTAB的方法提取剩余的较小部分。通过测试几种大小不同的DNA片段,可以测量DNA提取方法在PCR扩增成功率方面的性能。此外,为了估计DNA片段化水平,使用了自动芯片上电泳系统。
结果:结果表明,DNA纯度和靶基因区域的长度是驱动成功PCR反应的基本因素。相反,DNA产量和标本年龄似乎不太相关。通过这项研究,我还提出了一种基于CTAB的优化方法,该方法可以有效抑制植物标本室DNA中的抑制剂。可以认为该方法是基于列的技术的更便宜的替代方法,对处理大量样品特别有用。这项研究的结果证实了以前的报道,并有助于填补分子分析中现有的空白,其中涉及使用植物标本室收集的苔藓。

+1
+2
100研值 100研值 ¥99课程
检索文献一次
下载文献一次

去下载>

成功解锁2个技能,为你点赞

《SCI写作十大必备语法》
解决你的SCI语法难题!

技能熟练度+1

视频课《玩转文献检索》
让你成为检索达人!

恭喜完成新手挑战

手机微信扫一扫,添加好友领取

免费领《Endnote文献管理工具+教程》

微信扫码, 免费领取

手机登录

获取验证码
登录