Suppr超能文献

一种用于高效可靠地提取膳食样本DNA的自动化方案的评估。

Evaluation of an automated protocol for efficient and reliable DNA extraction of dietary samples.

作者信息

Wallinger Corinna, Staudacher Karin, Sint Daniela, Thalinger Bettina, Oehm Johannes, Juen Anita, Traugott Michael

机构信息

Mountain Agriculture Research Unit Institute of Ecology University of Innsbruck Innsbruck Austria.

出版信息

Ecol Evol. 2017 Jul 7;7(16):6382-6389. doi: 10.1002/ece3.3197. eCollection 2017 Aug.

Abstract

Molecular techniques have become an important tool to empirically assess feeding interactions. The increased usage of next-generation sequencing approaches has stressed the need of fast DNA extraction that does not compromise DNA quality. Dietary samples here pose a particular challenge, as these demand high-quality DNA extraction procedures for obtaining the minute quantities of short-fragmented food DNA. Automatic high-throughput procedures significantly decrease time and costs and allow for standardization of extracting total DNA. However, these approaches have not yet been evaluated for dietary samples. We tested the efficiency of an automatic DNA extraction platform and a traditional CTAB protocol, employing a variety of dietary samples including invertebrate whole-body extracts as well as invertebrate and vertebrate gut content samples and feces. Extraction efficacy was quantified using the proportions of successful PCR amplifications of both total and prey DNA, and cost was estimated in terms of time and material expense. For extraction of total DNA, the automated platform performed better for both invertebrate and vertebrate samples. This was also true for prey detection in vertebrate samples. For the dietary analysis in invertebrates, there is still room for improvement when using the high-throughput system for optimal DNA yields. Overall, the automated DNA extraction system turned out as a promising alternative to labor-intensive, low-throughput manual extraction methods such as CTAB. It is opening up the opportunity for an extensive use of this cost-efficient and innovative methodology at low contamination risk also in trophic ecology.

摘要

分子技术已成为实证评估捕食关系的重要工具。新一代测序方法使用的增加凸显了快速DNA提取的必要性,且这种提取不能影响DNA质量。饮食样本在这里构成了一项特殊挑战,因为获取微量短片段食物DNA需要高质量的DNA提取程序。自动高通量程序显著减少了时间和成本,并使总DNA提取标准化成为可能。然而,这些方法尚未针对饮食样本进行评估。我们测试了一种自动DNA提取平台和传统CTAB方案的效率,使用了各种饮食样本,包括无脊椎动物全身提取物以及无脊椎动物和脊椎动物的肠道内容物样本及粪便。提取效率通过总DNA和猎物DNA成功PCR扩增的比例来量化,成本则根据时间和材料费用进行估算。对于总DNA的提取,自动平台在无脊椎动物和脊椎动物样本中表现更佳。在脊椎动物样本的猎物检测方面也是如此。对于无脊椎动物的饮食分析,在使用高通量系统以获得最佳DNA产量时仍有改进空间。总体而言,自动DNA提取系统成为了一种有前景的替代方法,可替代如CTAB这种劳动密集型、低通量的手动提取方法。它为在低污染风险的情况下,在营养生态学中广泛使用这种具有成本效益和创新性的方法开辟了机会。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e3b3/5574753/4598729ee549/ECE3-7-6382-g001.jpg

文献检索

告别复杂PubMed语法,用中文像聊天一样搜索,搜遍4000万医学文献。AI智能推荐,让科研检索更轻松。

立即免费搜索

文件翻译

保留排版,准确专业,支持PDF/Word/PPT等文件格式,支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述,25分钟生成高质量综述,智能提取关键信息,辅助科研写作。

立即免费体验