Nucleic Acid Extraction and Sequencing from Low-Biomass Synthetic Mars Analog Soils for<i>In Situ</i>Life Detection

Mojarro, Angel; Hachey, Julie; Bailey, Ryan C.; Brown, Mark J.; Doebler, Robert; Ruvkun, Gary; Zuber, M. T.; Carr, Christopher E.

doi:10.1089/ast.2018.1929

Cited by 20 publications

(15 citation statements)

References 101 publications

(140 reference statements)

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“…Detection of nucleic acid biomarkers in extraterrestrial environments, for example, Mars, could indicate whether a signal came from organisms with common ancestries to those on Earth (Mojarro et al, 2017;Pontefract et al, 2017). The abundance of nucleic acid building blocks produced in the cosmos may have pushed all life to use nucleic acids as genetic materials (Callahan et al, 2011); thus, using nucleic acids as extraterrestrial biomarkers may provide an unambiguous biosignature.…”

Section: Fig 6 Complex Linkages Of the C And Si Cyclesmentioning

confidence: 99%

Deciphering Biosignatures in Planetary Contexts

et al. 2019

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Microbial life permeates Earth's critical zone and has likely inhabited nearly all our planet's surface and near subsurface since before the beginning of the sedimentary rock record. Given the vast time that Earth has been teeming with life, do astrobiologists truly understand what geological features untouched by biological processes would look like? In the search for extraterrestrial life in the Universe, it is critical to determine what constitutes a biosignature across multiple scales, and how this compares with “abiosignatures” formed by nonliving processes. Developing standards for abiotic and biotic characteristics would provide quantitative metrics for comparison across different data types and observational time frames. The evidence for life detection falls into three categories of biosignatures: (1) substances, such as elemental abundances, isotopes, molecules, allotropes, enantiomers, minerals, and their associated properties; (2) objects that are physical features such as mats, fossils including trace-fossils and microbialites (stromatolites), and concretions; and (3) patterns, such as physical three-dimensional or conceptual n -dimensional relationships of physical or chemical phenomena, including patterns of intermolecular abundances of organic homologues, and patterns of stable isotopic abundances between and within compounds. Five key challenges that warrant future exploration by the astrobiology community include the following: (1) examining phenomena at the “right” spatial scales because biosignatures may elude us if not examined with the appropriate instrumentation or modeling approach at that specific scale; (2) identifying the precise context across multiple spatial and temporal scales to understand how tangible biosignatures may or may not be preserved; (3) increasing capability to mine big data sets to reveal relationships, for example, how Earth's mineral diversity may have evolved in conjunction with life; (4) leveraging cyberinfrastructure for data management of biosignature types, characteristics, and classifications; and (5) using three-dimensional to n -D representations of biotic and abiotic models overlain on multiple overlapping spatial and temporal relationships to provide new insights.

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Section: Fig 6 Complex Linkages Of the C And Si Cyclesmentioning

confidence: 99%

Deciphering Biosignatures in Planetary Contexts

et al. 2019

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“…These interactions are further exacerbated in low-biomass environments and by nanopore sequencing, which presently requires substantial (~ 1 µg) input DNA not likely to be acquired from recalcitrant soils (without amplification) due to limitations related to sequencing efficiency and nanopore longevity (Mojarro et al, 2018). Roughly five in a million nucleobases are sequenced (R9.4 flowcells and chemistry) while nanopores are expended before detection of sub-nanogram input DNA (Mojarro et al, 2017a(Mojarro et al, , 2018 without library preparation modifications.…”

Section: Introductionmentioning

confidence: 99%

Nucleic acid extraction and sequencing from low-biomass synthetic Mars analog soils for in situ life detection

Mojarro

Hachey

Bailey

et al. 2018

Preprint

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Recent studies regarding the origin of life and Mars-Earth meteorite transfer simulations suggest that biological informational polymers, such as nucleic acids (DNA and RNA), have the potential to provide unambiguous evidence of life on Mars. To this end, we are developing a metagenomicsbased life-detection instrument which integrates nucleic acid extraction and nanopore sequencing:The Search for Extra-Terrestrial Genomes (SETG). Our goal is to isolate and sequence nucleic acids from extant or preserved life on Mars in order to determine if a particular genetic sequence (1) is distantly-related to life on Earth indicating a shared-ancestry due to lithological exchange, or (2) is unrelated to life on Earth suggesting a convergent origin of life on Mars. In this study, we validate prior work on nucleic acid extraction from cells deposited in Mars analog soils down to microbial concentrations observed in the driest and coldest regions on Earth. In addition, we report low-input nanopore sequencing results equivalent to 1 ppb life-detection sensitivity achieved by employing carrier sequencing, a method of sequencing sub-nanogram DNA in the background of a genomic carrier.

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“…NASA's Wetlab-2 platform onboard the ISS used the microbeadbeating technique without extraction buffers (OmniLyse from ClaremontBio) during the cell lysing step to extract and purify RNA (Parra et al, 2017). The Search for Extra-Terrestrial Genomes (SETG) instrument uses a modified Claremont BioSolutions solid-phase Purelyse bacterial genomic DNA extraction kit to isolate DNA and RNA from solid and liquid samples that are mixed with a binding buffer (low pH with ethylenediaminetetraacetic acid, EDTA; Mojarro et al, 2019). The adsorbed polynucleotides are then eluted with a high pH buffer containing EDTA.…”

Section: Bead Beating For Cell Lysismentioning

confidence: 99%

In situ organic biosignature detection techniques for space applications

Abrahamsson

Kanik

2022

Front. Astron. Space Sci.

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The search for life in Solar System bodies such as Mars and Ocean Worlds (e.g., Europa and Enceladus) is an ongoing and high-priority endeavor in space science, even ∼ five decades after the first life detection mission at Mars performed by the twin Viking landers. However, the in situ detection of biosignatures remains highly challenging, both scientifically and technically. New instruments are being developed for detecting extinct or extant life on Mars and Ocean Worlds due to new technology and fabrication techniques. These instruments are becoming increasingly capable of both detecting and identifying in situ organic biosignatures that are indicative of life and will play a pivotal role in the search for evidence of life through robotic lander missions. This review article gives an overview of techniques used for space missions (gas chromatography, mass spectrometry, and spectroscopy), the further ongoing developments of these techniques, and ion mobility spectrometry. In addition, current developments of techniques used in the next-generation instruments for organic biosignature detection are reviewed; these include capillary electrophoresis, liquid chromatography, biosensors (primarily immunoassays), and nanopore sensing; whereas microscopy, biological assays, and isotope analysis are beyond the scope of this paper and are not covered.

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Nucleic Acid Extraction and Sequencing from Low-Biomass Synthetic Mars Analog Soils forIn SituLife Detection

Cited by 20 publications

References 101 publications

Deciphering Biosignatures in Planetary Contexts

Deciphering Biosignatures in Planetary Contexts

Nucleic acid extraction and sequencing from low-biomass synthetic Mars analog soils for in situ life detection

In situ organic biosignature detection techniques for space applications

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