Real-Time DNA Sequencing in the Antarctic Dry Valleys Using the Oxford Nanopore Sequencer.
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.
Department of Biology, Georgetown University, Washington, DC 20057, USA;; Science, Technology, and International Affairs Program, Georgetown University, Washington, DC 20057, USA.
Sarah S
SS
Johnson
Department of Biology, Georgetown University, Washington, DC 20057, USA.
Elena
E
Zaikova
Georgetown University Medical Center, Washington, DC 20057, USA; and.
David S
DS
Goerlitz
Department of Biology, Georgetown University, Washington, DC 20057, USA.
Yu
Y
Bai
Advanced Genomics Lab, University of Vermont Cancer Center, Burlington, Vermont 05405, USA.
Scott W
SW
Tighe
D000864:Antarctic Regions; D010582:Bacteriophage lambda / Q000235:genetics; D003889:Desert Climate; D004783:Environmental Microbiology; D059014:High-Throughput Nucleotide Sequencing / Q000295:instrumentation*; D058889:Molecular Typing / Q000295:instrumentation*; D012015:Reference Standards; D017422:Sequence Analysis, DNA / Q000295:instrumentation*
Antarctica; MinION; Nanopore sequencing; extremophiles