CCPortal
DOI10.1371/journal.pone.0051273
Evaluating Ethanol-based Sample Preservation to Facilitate Use of DNA Barcoding in Routine Freshwater Biomonitoring Programs Using Benthic Macroinvertebrates
Stein, Eric D.1; White, Bryan P.1; Mazor, Raphael D.1; Miller, Peter E.2; Pilgrim, Erik M.3
发表日期2013-01-04
ISSN1932-6203
卷号8期号:1
英文摘要

Molecular methods, such as DNA barcoding, have the potential to enhance biomonitoring programs worldwide. Altering routinely used sample preservation methods to protect DNA from degradation may pose a potential impediment to application of DNA barcoding and metagenomics for biomonitoring using benthic macroinvertebrates. Using higher volumes or concentrations of ethanol, requirements for shorter holding times, or the need to include additional filtering may increase cost and logistical constraints to existing biomonitoring programs. To address this issue we evaluated the efficacy of various ethanol-based sample preservation methods at maintaining DNA integrity. We evaluated a series of methods that were minimally modified from typical field protocols in order to identify an approach that can be readily incorporated into existing monitoring programs. Benthic macroinvertebrates were collected from a minimally disturbed stream in southern California, USA and subjected to one of six preservation treatments. Ten individuals from five taxa were selected from each treatment and processed to produce DNA barcodes from the mitochondrial gene cytochrome c oxidase I (COI). On average, we obtained successful COI sequences (i.e. either full or partial barcodes) for between 93-99% of all specimens across all six treatments. As long as samples were initially preserved in 95% ethanol, successful sequencing of COI barcodes was not affected by a low dilution ratio of 2: 1, transfer to 70% ethanol, presence of abundant organic matter, or holding times of up to six months. Barcoding success varied by taxa, with Leptohyphidae (Ephemeroptera) producing the lowest barcode success rate, most likely due to poor PCR primer efficiency. Differential barcoding success rates have the potential to introduce spurious results. However, routine preservation methods can largely be used without adverse effects on DNA integrity.


语种英语
WOS记录号WOS:000313670100003
来源期刊PLOS ONE
来源机构美国环保署
文献类型期刊论文
条目标识符http://gcip.llas.ac.cn/handle/2XKMVOVA/59342
作者单位1.So Calif Coastal Water Res Project, Costa Mesa, CA USA;
2.Univ Guelph, Canadian Ctr DNA Barcoding, Biodivers Inst Ontario, Guelph, ON N1G 2W1, Canada;
3.US EPA, Natl Exposure Res Lab, Cincinnati, OH 45268 USA
推荐引用方式
GB/T 7714
Stein, Eric D.,White, Bryan P.,Mazor, Raphael D.,et al. Evaluating Ethanol-based Sample Preservation to Facilitate Use of DNA Barcoding in Routine Freshwater Biomonitoring Programs Using Benthic Macroinvertebrates[J]. 美国环保署,2013,8(1).
APA Stein, Eric D.,White, Bryan P.,Mazor, Raphael D.,Miller, Peter E.,&Pilgrim, Erik M..(2013).Evaluating Ethanol-based Sample Preservation to Facilitate Use of DNA Barcoding in Routine Freshwater Biomonitoring Programs Using Benthic Macroinvertebrates.PLOS ONE,8(1).
MLA Stein, Eric D.,et al."Evaluating Ethanol-based Sample Preservation to Facilitate Use of DNA Barcoding in Routine Freshwater Biomonitoring Programs Using Benthic Macroinvertebrates".PLOS ONE 8.1(2013).
条目包含的文件
条目无相关文件。
个性服务
推荐该条目
保存到收藏夹
导出为Endnote文件
谷歌学术
谷歌学术中相似的文章
[Stein, Eric D.]的文章
[White, Bryan P.]的文章
[Mazor, Raphael D.]的文章
百度学术
百度学术中相似的文章
[Stein, Eric D.]的文章
[White, Bryan P.]的文章
[Mazor, Raphael D.]的文章
必应学术
必应学术中相似的文章
[Stein, Eric D.]的文章
[White, Bryan P.]的文章
[Mazor, Raphael D.]的文章
相关权益政策
暂无数据
收藏/分享

除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。