Abstract
Rapid DNA sequencing and analysis has been a long-sought goal in remote research and point-of-care medicine. In microgravity, DNA sequencing can facilitate novel astrobiological research and close monitoring of crew health, but spaceflight places stringent restrictions on the mass and volume of instruments, crew operation time, and instrument functionality. The recent emergence of portable, nanopore-based tools with streamlined sample preparation protocols finally enables DNA sequencing on missions in microgravity. As a first step toward sequencing in space and aboard the International Space Station (ISS), we tested the Oxford Nanopore Technologies MinION during a parabolic flight to understand the effects of variable gravity on the instrument and data. In a successful proof-of-principle experiment, we found that the instrument generated DNA reads over the course of the flight, including the first ever sequenced in microgravity, and additional reads measured after the flight concluded its parabolas. Here we detail modifications to the sample-loading procedures to facilitate nanopore sequencing aboard the ISS and in other microgravity environments. We also evaluate existing analysis methods and outline two new approaches, the first based on a wave-fingerprint method and the second on entropy signal mapping. Computationally light analysis methods offer the potential for in situ species identification, but are limited by the error profiles (stays, skips, and mismatches) of older nanopore data. Higher accuracies attainable with modified sample processing methods and the latest version of flow cells will further enable the use of nanopore sequencers for diagnostics and research in space.
Highlights
Remote molecular diagnostics on Earth and in space necessitate portable technologies
Expanding on a proof-of-principle study regarding liquid handling in microgravity,[17] we demonstrated the possibility of performing a genomics experiment in space using the MinION sequencer
We did not observe any bubbles in the flow cell pre-flight, and we speculate that gravity or pressure changes may have contributed to the formation of the bubble
Summary
Remote molecular diagnostics on Earth and in space necessitate portable technologies. We initiated sequencing after sample loading while there were still ten parabolas remaining in the flight (~1 min each, including 30 s of microgravity) and continued sequencing through transport back to Johnson Space Center (Supplementary Figure 1).
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