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NASA NTRS1985

Native hole trap in bulk GaAs and its association with the double-charge state of the arsenic antisite defect

A dominant hole trap has been identified in p-type bulk GaAs employing deep level transient and photocapacitance spectroscopies. The trap is present at a concentration up to about 4 x 10 to the 16th per cu cm, and it has two charge states with energies 0.54 + or - 0.02 and 0.77 + or - 0.02 eV above the top of the valence band (at 77 K). From the upper level the trap can be photoexcited to a persistent metastable state just as the dominant midgap level, EL2. Impurity analysis and the photoionization characteristics rule out association of the trap with impurities Fe, Cu, or Mn. Taking into consideration theoretical results, it appears most likely that the two charge states of the trap are the single and double donor levels of the arsenic antisite As(Ga) defect.

Lagowski, J.↗

NASA NTRS1974

Evaluation of ERTS-1 data applications to geologic mapping, structural analysis and mineral resource inventory of South America with special emphasis on the Andes Mountain region

The author has identified the following significant results. A color composite of image E1010-14035, dated 2 August 1972, covers the west central Bolivian Altiplano near Salar de Coipasa. It clearly shows the distribution of surface water and scant patches of vegetation. The Salar de Coipasa is the largest body of water in the area, about 130 sq km of dark blue fresh water. A lighter blue area south of the lake suggests a thin cover of highly saline water superposed on salt beds. The scattered vegetation patches are presumed to be native grasses, lichens, and possibly Indian potato and maiz areas. A detailed study has been made of the scene which provides 12 different interpretive overlays including geology, volcanology, soils, hydrology, and relative permeability. It was found that color composites provide at least 40% more information that do black and white renditions. An excellent example of change detection was provided by image E1244-14051, dated 24 March 1973. Water in the Salar de Coipasa had more than doubled as a result of the rains of the Bolivian winter, which generally occur in the February-March period. The Salars are excellent and highly sensitive moisture indicators in this highly arid region.

Carter, W. D.↗

NASA NTRS2002

Crystallization and Preliminary X-ray Analysis of Der f 2, a Potent Allergen Derived from the House Dust Mite

Although a number of allergens have been identified and isolated, the underlying molecular basis for the potent immune response is poorly understood. House dust mites (Dermatophugoides sp.) are particularly ubiquitous contributors to atopy in developed countries. The rhinitis, dermatitis, and asthma associated with allergic reactions to these arthropods are often caused by relatively small (125-129 amino acids) mite proteins of unclear biological function. Der f 2, a major allergen from the mite Dermatophagoides farinae, has been recombinantly expressed and characterized. The Der f 2 protein has been crystallized in our laboratory and a native data set collected at a synchrotron source. The crystals belong to the orthorhombic space group I422 with unit cell parameters of a = 95.2 Angstroms, b = 95.2 Angstroms, and c = 103.3 Angstroms. An essentially complete (97.2%) data set has been collected to 2.4 Angstroms. Attempts to solve the crystal structure of Der f 2 by molecular replacement using the available NMR coordinates for either Der f 2 or Der p 2 (the homologous protein from D. pterovssinus) failed to reveal a creditable solution.

Roeber, Dana↗

NASA NTRS2018

File Specification for M2AMIP Products: Version Number - 1.0

The Modern-Era Retrospective analysis for Research and Applications, Version 2 (MERRA-2) is an atmospheric reanalysis computed with the Goddard Earth Observing (EOS) System, Version 5.12.4 (GEOS) data assimilation system (Gelaro et al., 2017). To supplement the reanalysis, the GEOS General Circulation Model (GCM) used in MERRA-2 has been used to generate a 10-member ensemble of simulations, configured following the convention of the Atmospheric Model Intercomparison Project (AMIP; Gates et al., 1992). Each ensemble member was initialized using meteorological fields from a different date in November 1979. The AMIP simulations used the sea-surface temperature (SST) and sea-ice boundary conditions that were used in MERRA-2 (Bosilovich et al., 2016). This 10-member ensemble of AMIP simulations, denoted M2AMIP, is available for download in a group of self-describing files, which are documented in this office note. All data collections are provided on the same horizontal grid as MERRA-2. This grid has 576 points in the longitudinal direction and 361 points in the latitudinal direction, corresponding to a resolution of 0.625 degrees by 0.5 degrees. Although data collections are available at this grid, all fields are computed on a cubed-sphere grid with an approximate resolution of 50 km by 50 km and are then spatially interpolated to the latitude-longitude grid. There are no changes in the vertical grids used: variables are provided on either the native vertical grid of 72 model layers, or interpolated to 42 standard pressure levels. Unlike MERRA, no data collections are available at the vertical layer edges. More details on the grid are provided in Section 4. MERRA-2 introduced observation-based precipitation forcing for the land surface parameterization and the corresponding variable PRECTOTCORR in the MERRA-2 FLX (surface turbulent fluxes and related quantities) and LFO (land-surface forcing) collections (see Section 6; Reichle et al., 2017). While this variable is still available for M2AMIP, there was no observation-based forcing, making the value identical to the model derived precipitation, PRECTOT. Similarly, without data assimilation, the values for the analysis increments, D*DTANA, in the tendency and vertically integrated file collections are zero. The M2AMIP data are available for download online through the NASA Center for Climate Simulation (NCCS) DataPortal (https://portal.nccs.nasa.gov/datashare/gmao_m2amip/). Data are arranged in subdirectories based on ensemble member, followed by year and month. Control files that are compatible with the Grid Analysis and Display System (GrADS) are available in the ctl_daily and ctl_monthly directories for the hourly, three hourly, and monthly mean data. Control files for the monthly mean diurnal cycle can be found in the ctl_diurnal subdirectory within the directory for each individual ensemble member.

M2AMIP↗

NASA NTRS2004

Microbial structural diversity estimated by dilution-extinction of phenotypic traits and T-RFLP analysis along a land-use intensification gradient

The present work tested whether the relationship between functional traits and inoculum density reflected structural diversity in bacterial communities from a land-use intensification gradient applying a mathematical model. Terminal restriction fragment length polymorphism (T-RFLP) analysis was also performed to provide an independent assessment of species richness. Successive 10-fold dilutions of a soil suspension were inoculated onto Biolog GN(R) microplates. Soil bacterial density was determined by total cell and plate counts. The relationship between phenotypic traits and inoculum density fit the model, allowing the estimation of maximal phenotypic potential (Rmax) and inoculum density (KI) at which Rmax will be half-reduced. Though Rmax decreased with time elapsed since clearing of native vegetation, KI remained high in two of the disturbed sites. The genetic pool of bacterial community did not experience a significant reduction, but the active fraction responding in the Biolog assay was adversely affected, suggesting a reduction in the functional potential. c2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

NASA Center KSC↗

NASA NTRS2013

Evidence for Perchlorates and the Origin of Chlorinated Hydrocarbons Detected by SAM at the Rocknest Aeolian Deposit in Gale Crater

A single scoop of the Rocknest aeolian deposit was sieved (less than 150 micrometers), and four separate sample portions, each with a mass of approximately 50 mg, were delivered to individual cups inside the Sample Analysis at Mars (SAM) instrument by the Mars Science Laboratory rover's sample acquisition system. The samples were analyzed separately by the SAM pyrolysis evolved gas and gas chromatograph mass spectrometer analysis modes. Several chlorinated hydrocarbons including chloromethane, dichloromethane, trichloromethane, a chloromethylpropene, and chlorobenzene were identified by SAM above background levels with abundances of approximately 0.01 to 2.3 nmol. The evolution of the chloromethanes observed during pyrolysis is coincident with the increase in O2 released from the Rocknest sample and the decomposition of a product of N-methyl-N-(tert-butyldimethylsilyl)-trifluoroacetamide (MTBSTFA), a chemical whose vapors were released from a derivatization cup inside SAM. The best candidate for the oxychlorine compounds in Rocknest is a hydrated calcium perchlorate (Ca(ClO4)2·nH2O), based on the temperature release of O2 that correlates with the release of the chlorinated hydrocarbons measured by SAM, although other chlorine-bearing phases are being considered. Laboratory analog experiments suggest that the reaction of Martian chlorine from perchlorate decomposition with terrestrial organic carbon from MTBSTFA during pyrolysis can explain the presence of three chloromethanes and a chloromethylpropene detected by SAM. Chlorobenzene may be attributed to reactions of Martian chlorine released during pyrolysis with terrestrial benzene or toluene derived from 2,6-diphenylphenylene oxide (Tenax) on the SAM hydrocarbon trap. At this time we do not have definitive evidence to support a nonterrestrial carbon source for these chlorinated hydrocarbons, nor do we exclude the possibility that future SAM analyses will reveal the presence of organic compounds native to the Martian regolith.

sample analysis at mars↗

NASA NTRS2017

A Dose of Reality: Radiation Analysis for Realistic Human Spacecraft

INTRODUCTION As with most computational analyses, a tradeoff exists between problem complexity, resource availability and response accuracy when modeling radiation transport from the source to a detector. The largest amount of analyst time for setting up an analysis is often spent ensuring that any simplifications made have minimal impact on the results. The vehicle shield geometry of interest is typically simplified from the original CAD design in order to reduce computation time, but this simplification requires the analyst to "re-draw" the geometry with a limited set of volumes in order to accommodate a specific radiation transport software package. The resulting low-fidelity geometry model cannot be shared with or compared to other radiation transport software packages, and the process can be error prone with increased model complexity. The work presented here demonstrates the use of the DAGMC (Direct Accelerated Geometry for Monte Carlo) Toolkit from the University of Wisconsin, to model the impacts of several space radiation sources on a CAD drawing of the US Lab module. METHODS The DAGMC toolkit workflow begins with the export of an existing CAD geometry from the native CAD to the ACIS format. The ACIS format file is then cleaned using SpaceClaim to remove small holes and component overlaps. Metadata is then assigned to the cleaned geometry file using CUBIT/Trelis from csimsoft (Registered Trademark). The DAGMC plugin script removes duplicate shared surfaces, facets the geometry to a specified tolerance, and ensures that the faceted geometry is water tight. This step also writes the material and scoring information to a standard input file format that the analyst can alter as desired prior to running the radiation transport program. The scoring results can be transformed, via python script, into a 3D format that is viewable in a standard graphics program. RESULTS The CAD model of the US Lab module of the International Space Station, inclusive of all the racks and components, was simplified to remove holes and volume overlaps. Problematic features within the drawing were also removed or repaired to prevent runtime issues. The cleaned drawing was then run through the DAGMC workflow to prepare for analysis. Pilot tests modeling transport of 1GeV proton and 800MeV/A oxygen sources show that reasonable results are converged upon in an acceptable amount of overall computation time from drawing preparation to data analysis. The FLUKA radiation transport code will next be used to model both a GCR and a trapped radiation source. These results will then be compared with measurements that have been made by the radiation instrumentation deployed inside the US Lab module. DISCUSSION Early analyses have indicated that the DAGMC workflow is a promising toolkit for running vehicle geometries of interest to NASA through multiple radiation transport codes. In addition, recent work has shown that a realistic human phantom, provided via a subcontract with the University of Florida, can be placed inside any vehicle geometry for a combinatorial analysis. This added functionality gives the user the ability to score various parameters at the organ level, and the results can then be used as input for cancer risk models.

Barzilla, J. E.↗

NASA NTRS2019

Verification Benchmarks and Validation of MAT261 for High Energy Dynamic Impact Applications

Advanced computational methods for composite structures can enable smarter testing and potentially certification by analysis. These methods, known as progressive damage and failure analysis (PDFA) methods, can predict lamina level mechanics including failure and subsequent propagation. When introduced into a commercial off the shelf finite element solver such as ABAQUS, LS-DYNA, or ANSYS, it enables end users to effectively exercise PDFA methods; however, it requires an understanding of the assumed mechanics and underlying coded response. In this paper, verification benchmarks developed under the NASA Advanced Composite Consortium for a class of PDFA methods known as continuum damage mechanics approaches are applied to the native LS-DYNA material model MAT261. As a result of the verification, an instability was discovered and subsequently corrected with an engineering solution. The results of the preliminary MAT261 simulations for high energy dynamic impact and the engineering solution are compared and a discussion about the significance is presented.

Molitor, Matthew↗

NASA NTRS2009

Synthesis, Decomposition and Characterization of Fe and Ni Sulfides and Fe and CO Nanoparticles for Aerospace Applications

We describe several related studies where simple iron, nickel, and cobalt complexes were prepared, decomposed, and characterized for aeronautics (Fischer-Tropsch catalysts) and space (high-fidelity lunar regolith simulant additives) applications. We describe the synthesis and decomposition of several new nickel dithiocarbamate complexes. Decomposition resulted in a somewhat complicated product mix with NiS predominating. The thermogravimetric analysis of fifteen tris(diorganodithiocarbamato)iron(III) has been investigated. Each undergoes substantial mass loss upon pyrolysis in a nitrogen atmosphere between 195 and 370 C, with major mass losses occurring between 279 and 324 C. Steric repulsion between organic substituents generally decreased the decomposition temperature. The product of the pyrolysis was not well defined, but usually consistent with being either FeS or Fe2S3 or a combination of these. Iron nanoparticles were grown in a silica matrix with a long-term goal of introducing native iron into a commercial lunar dust simulant in order to more closely simulate actual lunar regolith. This was also one goal of the iron and nickel sulfide studies. Finally, cobalt nanoparticle synthesis is being studied in order to develop alternatives to crude processing of cobalt salts with ceramic supports for Fischer-Tropsch synthesis.

Cowen, Jonathan E.↗

NASA NTRS2017

Giovanni in the Cloud: Earth Science Data Exploration in Amazon Web Services

Giovanni is an exploration tool at the NASA Goddard Earth Sciences Data Information Services Center (GES DISC), providing 22 analysis and visualization services for over 1600 Earth Science data variables. Owing to its popularity, Giovanni has experienced a consistent growth in overall demand, with periodic usage spikes attributed to trainings by education organizations, extensive data analysis in response to natural disasters, preparations for science meetings, etc. Furthermore, the new generation of spaceborne sensors and high resolution models have resulted in an exponential growth in data volume with data distributed across the traditional boundaries of data centers. Seamless exploration of data (without users having to worry about data center boundaries) has been a key recommendation of the GES DISC User Working Group. These factors have required new strategies for delivering acceptable performance. The cloud-based Giovanni, built on Amazon Web Services (AWS), evaluates (1) AWS native solutions to provide a scalable, serverless architecture; (2) open standards for data storage in the Cloud; (3) a cost model for operations; and (4) end-user performance. Our preliminary findings indicate that the use of serverless architecture has a potential to significantly reduce development and operational cost of Giovanni. The combination of using AWS managed services, storage of data in open standards, and schema-on-read data access strategy simplifies data access and analytics, in addition to making data more accessible to the end users of Giovanni through popular programming languages.

Giovanni↗

NASA NTRSDate not supplied

ELM: Europa Luminescence Microscope

The Europa Luminescence Microscope (ELM) is an automated fluorescence and bright-field microscope designed to meet key objectives defined in the 2016 NASA Europa Lander Study Report, including the identification and characterization of morphological biosignatures. ELM’s heritage stems from a 2U cubesat fluorescence microscope, the Fluorescence Analysis for In situ Research imager, designed and built at NASA Ames Research Center, for the autonomous study of microbial biology in low Earth orbit. For ELM implementation, a sample is autonomously manipulated with a microfluidic system using in-line 10, 1.0, and 0.2 μm pore-size filters to capture successively smaller particles for imaging. For bright-field imaging, ELM uses deep-ultraviolet, ultraviolet and visible light to image organic and inorganic structures with submicron resolution. The ability to detect biosignatures as small as 0.2 μm in size is achieved by imaging native fluorescence and using fluorescence microscopy stains to identify key structural and functional indicators of microbial life (proteins, lipids, nucleic acids). For fluorescence imaging, ELM uses 265, 370, 470, and 530 nm LEDs with five emission bands. The use of multiple excitation and emission wavelengths for native fluorescence imaging enables the detection of a wide range of molecular species and their rough classification. Excitation at 265 nm allows for the detection of smaller polycyclic aromatic hydrocarbons (PAH), aromatic amino acids, and proteins with little to no interference from mineral fluorescence, given proper emission band selection. 370 and 470 nm light excites increasingly larger PAH structures and larger aromatic biomolecules that may be present (e.g., protective pigments). Similarly, inorganic fluorescence can be characterized and separated from organic fluorescence, allowing the recognition and in some cases classification, of minerals and other abiotic particles. ELM is based upon work supported by the NASA COLDTech and ICEE-2 programs.

Richard C Quinn↗

NASA NTRS2015

Mapping Species Composition of Forests and Tree Plantations in Northeastern Costa Rica with an Integration of Hyperspectral and Multitemporal Landsat Imagery

An efficient means to map tree plantations is needed to detect tropical land use change and evaluate reforestation projects. To analyze recent tree plantation expansion in northeastern Costa Rica, we examined the potential of combining moderate-resolution hyperspectral imagery (2005 HyMap mosaic) with multitemporal, multispectral data (Landsat) to accurately classify (1) general forest types and (2) tree plantations by species composition. Following a linear discriminant analysis to reduce data dimensionality, we compared four Random Forest classification models: hyperspectral data (HD) alone; HD plus interannual spectral metrics; HD plus a multitemporal forest regrowth classification; and all three models combined. The fourth, combined model achieved overall accuracy of 88.5%. Adding multitemporal data significantly improved classification accuracy (p less than 0.0001) of all forest types, although the effect on tree plantation accuracy was modest. The hyperspectral data alone classified six species of tree plantations with 75% to 93% producer's accuracy; adding multitemporal spectral data increased accuracy only for two species with dense canopies. Non-native tree species had higher classification accuracy overall and made up the majority of tree plantations in this landscape. Our results indicate that combining occasionally acquired hyperspectral data with widely available multitemporal satellite imagery enhances mapping and monitoring of reforestation in tropical landscapes.

hyperspectral fusion↗

NASA NTRS2020

DEVELOP Poster: Medicine Bow Disasters

The Medicine Bow National Forest (MBNF) consists of approximately 1,383,790 acres of forested land, grassland, and sagebrush steppe in southeastern Wyoming. Cheatgrass (Bromus tectorum), an invasive plant species in the Western US, occurs in the grasslands throughout MBNF. Cheatgrass is known to rapidly colonize disturbed sites and dramatically alter historic fire regimes and nutrient/water dynamics as well as outcompete native plant species that are important forage for mule deer (Odocoileus hemionus) and elk (Cervus canadensis). In 2012, the Squirrel Creek Fire burned approximately 10,587 acres of land within MBNF, exacerbating the spread of cheatgrass. In 2015, the Wyoming Ecological Forecasting DEVELOP team identified areas of high cheatgrass abundance within the fire boundary in order to guide US Forest Service (USFS) herbicide spraying efforts to reduce cheatgrass in 2016. This research used Landsat 8 Operational Land Imager (OLI) and Sentinel-2 MultiSpectral Instrument (MSI) data to create a 2019 probabilistic cheatgrass occurrence map. This map allowed an analysis of the effectiveness of aerial spraying to inform future land management techniques for the USFS. Based on the results of the Generalized Linear Model, we found that treated areas decreased in cheatgrass cover by 36% while untreated areas increased in cheatgrass cover by 6%, suggesting that herbicide treatment has been effective.

Disasters↗

NASA NTRSDate not supplied

Europa Luminescence Microscope

The Europa Luminescence Microscope (ELM) is an automated fluorescence and dark-field mi-croscope designed to meet key objectives defined in the NASA Europa Lander Study Report, includ-ing the identification and characterization of morphological biosignatures. ELM’s heritage stems from a 2U cubesat fluorescence microscope, the Fluorescence Analysis for In-situ Research (FLAIR) im-ager, designed and built at NASA Ames Research Center, for the autonomous study of microbial bi-ology in low Earth orbit. For the ELM implementation, a sample is autonomously manipulated with a microfluidic system using in-line filter sets to capture successively smaller particles on 10, 1.0, and 0.1 µm pore-size filters for imaging. For darkfield imaging, ELM uses ultraviolet and visible light to image organic and inorganic structures with submicron resolution. The ability to detect structural and chemical biosignatures as small as 0.2 µm in size is achieved by imaging native fluorescence and us-ing fluorescence microscopy stains to identify key molecular and structural indicators of microbial life (proteins, lipids, nucleic acids). To excite fluorescence, ELM uses LEDs with wavelengths centered near 265, 370, 470, and 530 nm and five emission bands. The use of multiple excitation and emission wavelengths for native fluorescence imaging not only enables the detection of different molecular species, but also their rough classification. Excitation at 265 nm allows for the detection of smaller polyaromatic hydrocarbons (PAHs; 1-5 rings), aromatic amino acids, and proteins with little to no interference from mineral fluorescence, given proper emission band selection. 370 and 470 nm light excites increasingly larger PAH structures (e.g., coronene) and larger aromatic biomolecules that may be present (e.g., protective pigments). Similarly, inorganic fluorescence can be characterized and sep-arated from organic fluorescence, allowing the recognition and in some cases classification, of miner-als and other abiotic particles. ELM is based upon work supported by the NASA COLDTech and ICEE-2 programs.

Microscope↗

NASA NTRS1990

Robotic lunar surface operations: Engineering analysis for the design, emplacement, checkout and performance of robotic lunar surface systems

The assembly, emplacement, checkout, operation, and maintenance of equipment on planetary surfaces are all part of expanding human presence out into the solar system. A single point design, a reference scenario, is presented for lunar base operations. An initial base, barely more than an output, which starts from nothing but then quickly grows to sustain people and produce rocket propellant. The study blended three efforts: conceptual design of all required surface systems; assessments of contemporary developments in robotics; and quantitative analyses of machine and human tasks, delivery and work schedules, and equipment reliability. What emerged was a new, integrated understanding of hot to make a lunar base happen. The overall goal of the concept developed was to maximize return, while minimizing cost and risk. The base concept uses solar power. Its primary industry is the production of liquid oxygen for propellant, which it extracts from native lunar regolith. Production supports four lander flights per year, and shuts down during the lunar nighttime while maintenance is performed.

Woodcock, Gordon R.↗

NASA NTRS1997

Multiple forms of ADP-glucose pyrophosphorylase from tomato fruit

ADP-glucose pyrophosphorylase (AGP) was purified from tomato (Lycopersicon esculentum Mill.) fruit to apparent homogeneity. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis the enzyme migrated as two close bands with molecular weights of 50,000 and 51,000. Two-dimensional polyacrylamide gel electrophoresis analysis of the purified enzyme, however, revealed at least five major protein spots that could be distinguished by their slight differences in net charge and molecular weight. Whereas all of the spots were recognized by the antiserum raised against tomato fruit AGP holoenzyme, only three of them reacted strongly with antiserum raised against the potato tuber AGP large subunit, and the other two spots (with lower molecular weights) reacted specifically with antisera raised against spinach leaf AGP holoenzyme and the potato tuber AGP small subunit. The results suggest the existence of at least three isoforms of the AGP large subunit and two isoforms of the small subunit in tomato fruit in vivo. The native molecular mass of the enzyme determined by gel filtration was 220 +/- 10 kD, indicating a tetrameric structure for AGP from tomato fruit. The purified enzyme is very sensitive to 3-phosphoglycerate/inorganic phosphate regulation.

NASA Discipline Life Support Systems↗

NASA NTRS2007

Improvements to NASA's Debris Assessment Software

NASA's Debris Assessment Software (DAS) has been substantially revised and expanded. DAS is designed to assist NASA programs in performing orbital debris assessments, as described in NASA s Guidelines and Assessment Procedures for Limiting Orbital Debris. The extensive upgrade of DAS was undertaken to reflect changes in the debris mitigation guidelines, to incorporate recommendations from DAS users, and to take advantage of recent software capabilities for greater user utility. DAS 2.0 includes an updated environment model and enhanced orbital propagators and reentry-survivability models. The ORDEM96 debris environment model has been replaced by ORDEM2000 in DAS 2.0, which is also designed to accept anticipated revisions to the environment definition. Numerous upgrades have also been applied to the assessment of human casualty potential due to reentering debris. Routines derived from the Object Reentry Survival Analysis Tool, Version 6 (ORSAT 6), determine which objects are assessed to survive reentry, and the resulting risk of human casualty is calculated directly based upon the orbital inclination and a future world population database. When evaluating reentry risks, the user may enter up to 200 unique hardware components for each launched object, in up to four nested levels. This last feature allows the software to more accurately model components that are exposed below the initial breakup altitude. The new DAS 2.0 provides an updated set of tools for users to assess their mission s compliance with the NASA Safety Standard and does so with a clear and easy-to-understand interface. The new native Microsoft Windows graphical user interface (GUI) is a vast improvement over the previous DOS-based interface. In the new version, functions are more-clearly laid out, and the GUI includes the standard Windows-style Help functions. The underlying routines within the DAS code are also improved.

Opiela, J.↗

NASA NTRS2017

Novel End-to-End Molecular Biology Approach for Direct Nanopore 1D cDNA Sequencing of Reverse Transcribed mRNAs Purified from Cell Cultures by the NASA ISS WetLab2 SPM

Continued space bioscience research onboard the International Space Station (ISS) and future long-duration flight missions to the Moon or Mars will require the ability to conduct on-orbit molecular analysis of biological samples independently from Earth. In the last year two new molecular analytic technologies have been installed and the technologies demonstrated onboard the ISS: The Sample Prep Module (SPM) WetLab-2 (WL2) qRT-PCR toolbox and the Oxford Nanopore MinIon Biomolecule Sequencer. Here we describe protocol development and integration into existing ISS technology for end-to-end on-orbit biological sample processing and molecular analysis with real time results generated utilizing only field offline analytic software. For this experiment we isolated primary cells from bone marrow flushes of wild type B6129SF2 mice (Jackson Labs) long bones. The cell isolate was then processed using the SPM to produce total 147nanograms of RNA. The total RNA was purified to only messenger RNA (mRNA) and transferred to Smartcycler Thermocycle ISS kit consumable tube using Eppendorf gel loading pipette tips for further processing. Complementary first strand cDNA was synthesized using OLIGO dT priming followed by addition of SuperScript II Reverse Transcriptase and thermal cycling as per manufacturers instruction. All thermal cycling was conducted using the ISS WetLab-2 Cephid Smarcycler real time thermal cycler. Our protocol takes advantage of mRNAs native poly(A) tail, synthesized in vivo to protect the mRNA from degradation by endonucleases, to eliminate end-prep for adapter ligation. The adapted library is purified using MyOne C1 Streptavidin beads before elution in buffer. The pre-sequencing library is diluted in the loading buffer and injected into the MinIon sample port, drawn into the nanopore window by capillary action, and sequenced using the MinKnown software with local basecalling. The sequencing read produced 34.5 million events and local basecalling produced 117,301 successful reads. NCBI Blast of the data for the mouse genome resulted in 2,462 successful nucleotide collection matches (gene sequences) exceeding 70 homology. These results demonstrate the viability of this novel flight ready end-to-end sample analytic methodology and provide a real time homolog for flight experimentation utilizing supply kits and technologies that have already been demonstrated on ISS.

MinIon↗