DNA Extraction and Preparation for a Sequencing Process from Low Biomass Samples (DEAP Seq PLS)

As we set our sights toward Mars, we must develop the means to perform environmental and medical monitoring in the spaceflight environment. Determining the sequence of DNA is a powerful molecular biology technique that enables medical diagnostics and the identification of microorganisms. A DNA sequencer is being tested on the ISS in 2016 by the proposal team (Biomolecule Sequencer Payload). Sequence-ready DNA, prepared on the ground, will be launched for this testing. The ability to perform inflight sample collection and DNA manipulations, required to generate sequence-ready DNA, from low biomass samples (few cells and/or free DNA, as is the case on the ISS and in planetary protection scenarios) is far from reality. Having the capability to preform inflight sequencing preparation could expand the capabilities of the sequencer to include microbial monitoring, monitoring changes in microbes and humans in response to spaceflight, addressing planetary protection concerns, and detecting DNA-based life elsewhere in the universe.

This project will develop a common process including: 1) optimal swab selection, 2) DNA extraction, 3) DNA manipulation, and 4) the preparation of sequence-ready DNA that is compatible with the sequencer being tested inflight, for cell and free DNA collection from high-fidelity, low biomass samples. The development of a sample collection and preparation process is essential for the sequencer to be used in a capacity beyond testing Earth-prepared DNA samples.