Johnson Sarah S, Zaikova Elena, Goerlitz David S, Bai Yu, Tighe Scott W
Department of Biology, Georgetown University, Washington, DC 20057, USA;; Science, Technology, and International Affairs Program, Georgetown University, Washington, DC 20057, USA.
Department of Biology, Georgetown University, Washington, DC 20057, USA.
J Biomol Tech. 2017 Apr;28(1):2-7. doi: 10.7171/jbt.17-2801-009. Epub 2017 Mar 22.
The ability to sequence DNA outside of the laboratory setting has enabled novel research questions to be addressed in the field in diverse areas, ranging from environmental microbiology to viral epidemics. Here, we demonstrate the application of offline DNA sequencing of environmental samples using a hand-held nanopore sequencer in a remote field location: the McMurdo Dry Valleys, Antarctica. Sequencing was performed using a MK1B MinION sequencer from Oxford Nanopore Technologies (ONT; Oxford, United Kingdom) that was equipped with software to operate without internet connectivity. One-direction (1D) genomic libraries were prepared using portable field techniques on DNA isolated from desiccated microbial mats. By adequately insulating the sequencer and laptop, it was possible to run the sequencing protocol for up to 2½ h under arduous conditions.
在实验室环境之外对DNA进行测序的能力,使得在从环境微生物学到病毒流行等不同领域中能够解决新的研究问题。在这里,我们展示了在偏远野外地点——南极洲麦克默多干谷,使用手持式纳米孔测序仪对环境样本进行离线DNA测序的应用。测序使用的是牛津纳米孔技术公司(ONT;英国牛津)的MK1B MinION测序仪,该测序仪配备了无需互联网连接即可运行的软件。使用便携式野外技术从干燥的微生物垫中分离出的DNA制备单向(1D)基因组文库。通过对测序仪和笔记本电脑进行充分绝缘,在艰苦条件下运行测序协议长达2个半小时成为可能。