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At least 613 records · Page 34

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NASA NTRS2011

Non-Contact Thermal Properties Measurement with Low-Power Laser and IR Camera System

As shown by the Phoenix Mars Lander's Thermal and Electrical Conductivity Probe (TECP), contact measurements of thermal conductivity and diffusivity (using a modified flux-plate or line-source heat-pulse method) are constrained by a number of factors. Robotic resources must be used to place the probe, making them unavailable for other operations for the duration of the measurement. The range of placement is also limited by mobility, particularly in the case of a lander. Placement is also subject to irregularities in contact quality, resulting in non-repeatable heat transfer to the material under test. Most important from a scientific perspective, the varieties of materials which can be measured are limited to unconsolidated or weakly-cohesive regolith materials, rocks, and ices being too hard for nominal insertion strengths. Accurately measuring thermal properties in the laboratory requires significant experimental finesse, involving sample preparation, controlled and repeatable procedures, and, practically, instrumentation much more voluminous than the sample being tested (heater plates, insulation, temperature sensors). Remote measurements (infrared images from orbiting spacecraft) can reveal composite properties like thermal inertia, but suffer both from a large footprint (low spatial resolution) and convolution of the thermal properties of a potentially layered medium. In situ measurement techniques (the Phoenix TECP is the only robotic measurement of thermal properties to date) suffer from problems of placement range, placement quality, occupation of robotic resources, and the ability to only measure materials of low mechanical strength. A spacecraft needs the ability to perform a non-contact thermal properties measurement in situ. Essential components include low power consumption, leveraging of existing or highly-developed flight technologies, and mechanical simplicity. This new in situ method, by virtue of its being non-contact, bypasses all of these problems. The use of photons to both excite and measure the thermal response of any surface material to a high resolution (estimated footprint = 10 square centimeters) is a generational leap in physical properties measurements. The proposed method consists of spot-heating the surface of a material with a low (less than 1 W) power laser. This produces a moderate (5-10 K) temperature increase in the material.

Hudson, Troy L.↗

DOE OSTI2017

Supercritical drying of cementitious materials

Techniques to characterize the microstructure of hydrated cement require dried materials. However, the microstructure of hydrated products is significantly altered by high capillary forces during drying when using the conventional drying methods. To avoid drying stresses when preparing samples, we have employed supercritical drying (SCD) which has been used for decades to prepare aerogels that undergo no shrinkage during drying, but has rarely been used for cementitious materials. The pore solution is first replaced with isopropanol, and then with trifluoromethane (R23). The temperature and pressure are raised above the critical point, where no menisci or capillary pressure can exist; therefore, the dried samples are free of artifacts created by stresses. Images from scanning electron microscope show less compact morphology for supercritically dried samples than that dried by conventional methods, while BET surface areas of SCD samples are very close to samples dried by the isopropanol replacement method. This can be explained by the fact that isopropanol and supercritical fluid enter the micropores and block them. The nature of the chemical interactions of isopropanol and R23 with cement pastes are still not clear, but no reaction products were identified in the present study.

36 MATERIALS SCIENCE↗

DOE OSTI2014

Characterization of the thermal conductivity of La 0.95 Sr 0.05 CoO 3 thermoelectric oxide nanofibers

Thermoelectric oxide nanofibers prepared by electrospinning are expected to have reduced thermal conductivity when compared to bulk samples. Measurements of nanofibers’ thermal conductivity is challenging since it involves sophisticated sample preparation methods. In this work, we present a novel method suitable for measurements of thermal conductivity of a single nanofiber. A microelectro-mechanical (MEMS) device has been designed and fabricated to perform thermal conductivity measurements on a single nanofiber. A special Si template was designed to collect and transfer individual nanofibers onto a MEMS device. Pt was deposited by a focused ion beam to reduce the effective length of a prepared nanofiber. La 0.95 Sr 0.05 CoO 3 nanofibers with diameters of 140 nm and 290 nm were studied and characterized using this approach at room temperature. Measured thermal conductivities yielded values of 0.7 W·m -1 ·K -1 and 2.1 W·m -1 ·K -1 , respectively. Our measurements in La 0.95 Sr 0.05 CoO 3 nanofibers confirmed that a decrease of linear dimensions has a profound effect on its thermal conductivity.

36 MATERIALS SCIENCE↗

DOE OSTI2018

Diagnosing and Correcting Mass Accuracy and Signal Intensity Error Due to Initial Ion Position Variations in a MALDI TOFMS

Frustrated by worse than expected error for both peak area and time-of-flight (TOF) in matrix assisted laser desorption ionization (MALDI) experiments using samples prepared by electrospray deposition, it was finally determined that there was a correlation between sample location on the target plate and the measured TOF/peak area. Variations in both TOF and peak area were found to be due to small differences in the initial position of ions formed in the source region of the TOF mass spectrometer. These differences arise largely from misalignment of the instrument sample stage, with a smaller contribution arising from the non-ideal shape of the target plates used. By physically measuring the target plates used and comparing TOF data collected from three different instruments, an estimate of the magnitude and direction of the sample stage misalignment was determined for each of the instruments. A correction method was developed to correct the TOFs and peak areas obtained for a given combination of target plate and instrument. Two correction factors are determined, one by initially collecting spectra from each sample position used and another by using spectra from a single position for each set of samples on a target plate. For TOF and mass values, use of the correction factor reduced the error by a factor of 4, with the relative standard deviation (RSD) of the corrected masses being reduced to 12-24 ppm. For the peak areas, the RSD was reduced from 28% to 16% for samples deposited twice onto two target plates over two days. .

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

DOE OSTI2018

Application of modern autoradiography to nuclear forensic analysis

Modern autoradiography techniques based on phosphorimaging technology using image plates (IPs) and digital scanning can identify heterogeneities in activity distributions and reveal material properties, serving to inform subsequent analyses. Here, we have adopted these advantages for applications in nuclear forensics, the technical analysis of radioactive or nuclear materials found outside of legal control to provide data related to provenance, production history, and trafficking route for the materials. IP autoradiography is a relatively simple, non-destructive method for sample characterization that records an image reflecting the relative intensity of alpha and beta emissions from a two-dimensional surface. Such data are complementary to information gathered from radiochemical characterization via bulk counting techniques, and can guide the application of other spatially resolved techniques such as scanning electron microscopy (SEM) and secondary ion mass spectrometry (SIMS). IP autoradiography can image large 2-dimenstional areas (up to 20 × 40 cm), with relatively low detection limits for actinides and other radioactive nuclides, and sensitivity to a wide dynamic range (10 5 ) of activity density in a single image. Distributions of radioactivity in nuclear materials can be generated with a spatial resolution of approximately 50 μm using IP autoradiography and digital scanning. While the finest grain silver halide films still provide the best possible resolution (down to ~10 μm), IP autoradiography has distinct practical advantages such as shorter exposure times, no chemical post-processing, reusability, rapid plate scanning, and automated image digitization. Sample preparation requirements are minimal, and the analytical method does not consume or alter the sample. These advantages make IP autoradiography ideal for routine screening of nuclear materials, and for the identification of areas of interest for subsequent micro-characterization methods. Here in this article we present a summary of our setup, as modified for nuclear forensic sample analysis and related research, and provide examples of data from select samples from the nuclear fuel cycle and historical nuclear test debris.

07 ISOTOPE AND RADIATION SOURCES↗

DOE OSTI2013

Tomographic investigation of fermi level pinning at focused ion beam milled semiconductor surfaces

Electron holography in the transmission electron microscope (TEM) offers the spatial and signal resolution for studying effects like Fermi level pinning or dopant concentration variations important for the design of modern electronic devices. To overcome the loss of information along the projection direction, surface effects, and surface damage due to TEM specimen preparation, we apply electron holographic tomography to analyze the 3D potential distribution of semiconductor samples prepared by focused-ion-beam. We observe mid-band gap pinning of the Fermi level at Si surfaces but valence band pinning at Ge surfaces. The pinning extends over tens of nanometers into the bulk.

36 MATERIALS SCIENCE↗

DOE OSTI2016

Study of mass attenuation coefficients and effective atomic numbers of bismuth-ground granulated blast furnace slag concretes

Five samples of Bismuth-Ground granulated blast furnace slag (Bi-GGBFS) concretes were prepared using composition (0.6 cement + x Bi{sub 2}O{sub 3} + (0.4-x) GGBFS, x = 0.05, 0.10, 0.15, 0.20 and 0.25) by keeping constant water (W) cement (C) ratio. Mass attenuation coefficients (μ{sub m}) of these prepared samples were calculated using a computer program winXCOM at different gamma ray energies, whereas effective atomic numbers (Z{sub eff}) is calculated using mathematical formulas. The radiation shielding properties of Bi-GGBFS concrete has been compared with standard radiation shielding concretes.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

DOE OSTI2021

X-ray microscopy enables multiscale high-resolution 3D imaging of plant cells, tissues, and organs

Capturing complete internal anatomies of plant organs and tissues within their relevant morphological context remains a key challenge in plant science. While plant growth and development are inherently multiscale, conventional light, fluorescence, and electron microscopy platforms are typically limited to imaging of plant microstructure from small flat samples that lack a direct spatial context to, and represent only a small portion of, the relevant plant macrostructures. We demonstrate technical advances with a lab-based X-ray microscope (XRM) that bridge the imaging gap by providing multiscale high-resolution three-dimensional (3D) volumes of intact plant samples from the cell to the whole plant level. Serial imaging of a single sample is shown to provide sub-micron 3D volumes co-registered with lower magnification scans for explicit contextual reference. High-quality 3D volume data from our enhanced methods facilitate sophisticated and effective computational segmentation. Advances in sample preparation make multimodal correlative imaging workflows possible, where a single resin-embedded plant sample is scanned via XRM to generate a 3D cell-level map, and then used to identify and zoom in on sub-cellular regions of interest for high-resolution scanning electron microscopy. In total, we present the methodologies for use of XRM in the multiscale and multimodal analysis of 3D plant features using numerous economically and scientifically important plant systems.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2012

Synthesis of mesoporous TS-1 using a hybrid SiO{sub 2}–TiO{sub 2} xerogel for catalytic oxidative desulfurization

Graphical abstract: Display Omitted Highlights: ► Meso-TS-1 catalyst was synthesized using a SiO{sub 2}–TiO{sub 2} xerogel with an organosilane precursor. ► Hierarchical pore structure was confirmed by characterization of the materials. ► Catalytic activity was tested using oxidative desulfurization of the model sulfur compounds. ► Meso-TS-1 demonstrated significantly improved catalytic activity than TS-1. -- Abstract: Mesoporous TS-1 (M-TS-1) was synthesized using a hybrid SiO{sub 2}–TiO{sub 2} xerogel combined with an organosilane precursor. Prepared samples were characterized by XRD, UV–vis spectroscopy, SEM, and N{sub 2} adsorption–desorption measurement. M-TS-1, prepared in 2 days, showed high crystallinity and the best textural properties among the samples. The N{sub 2} adsorption–desorption isotherms of M-TS-1 exhibited a hysteresis loop at pressure higher than P/P{sub 0} = 0.4, clearly indicating the existence of mesopores. M-TS-1 has significantly larger mesopore volume (0.48 cm{sup 3}/g) than that of conventional TS-1 (0.07 cm{sup 3}/g), and showed a narrow peak centered at ca. 6.3 nm. In the oxidative desulfurization reaction, M-TS-1 was more active than conventional TS-1 at the same Ti-loading; M-TS-1 produced a dibenzothiophene (DBT) conversion of 96%, whereas conventional TS-1 produced a final DBT conversion of 5.6% after a reaction time of 180 min. Oxidative desulfurization over TS-1 was influenced both by electron density and steric hindrance in the sulfur compounds tested.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2013

Rapid Quantitation of Ascorbic and Folic Acids in SRM 3280 Multivitamin/Multielement Tablets using Flow-Injection Tandem Mass Spectrometry

RATIONALE: Ascorbic acid (AA) and folic acid (FA) are water-soluble vitamins and are usually fortified in food and dietary supplements. For the safety of human health, proper intake of these vitamins is recommended. Improvement in the analysis time required for the quantitative determination of these vitamins in food and nutritional formulations is desired. METHODS: A simple and fast (~5 min) in-tube sample preparation was performed, independently for FA and AA, by mixing extraction solvent with a powdered sample aliquot followed by agitation, centrifugation, and filtration to recover an extract for analysis. Quantitative detection was achieved by flow-injection (1 L injection volume) electrospray ionization tandem mass spectrometry (ESI-MS/MS) in negative ion mode using the method of standard addition. RESULTS: Method of standard addition was employed for the quantitative estimation of each vitamin in a sample extract. At least 2 spiked and 1 non-spiked sample extract were injected in triplicate for each quantitative analysis. Given an injection-to-injection interval of approximately 2 min, about 18 min was required to complete the quantitative estimation of each vitamin. The concentration values obtained for the respective vitamins in the standard reference material (SRM) 3280 using this approach were within the statistical range of the certified values provided in the NIST Certificate of Analysis. The estimated limit of detections of FA and AA were 13 and 5.9 ng/g, respectively. CONCLUSIONS: Flow-injection ESI-MS/MS was successfully applied for the rapid quantitation of FA and AA in SRM 3280 multivitamin/multielement tablets.

ascorbic acid↗

DOE OSTI2012

Rapid determination of 226Ra in environmental samples

A new rapid method for the determination of {sup 228}Ra in natural water samples has been developed at the SRNL/EBL (Savannah River National Lab/ Environmental Bioassay Laboratory) that can be used for emergency response or routine samples. While gamma spectrometry can be employed with sufficient detection limits to determine {sup 228}Ra in solid samples (via {sup 228}Ac) , radiochemical methods that employ gas flow proportional counting techniques typically provide lower MDA (Minimal Detectable Activity) levels for the determination of {sup 228}Ra in water samples. Most radiochemical methods for {sup 228}Ra collect and purify {sup 228}Ra and allow for {sup 228}Ac daughter ingrowth for ~36 hours. In this new SRNL/EBL approach, {sup 228}Ac is collected and purified from the water sample without waiting to eliminate this delay. The sample preparation requires only about 4 hours so that {sup 228}Ra assay results on water samples can be achieved in < 6 hours. The method uses a rapid calcium carbonate precipitation enhanced with a small amount of phosphate added to enhance chemical yields (typically >90%), followed by rapid cation exchange removal of calcium. Lead, bismuth, uranium, thorium and protactinium isotopes are also removed by the cation exchange separation. {sup 228}Ac is eluted from the cation resin directly onto a DGA Resin cartridge attached to the bottom of the cation column to purify {sup 228}Ac. DGA Resin also removes lead and bismuth isotopes, along with Sr isotopes and {sup 90}Y. La is used to determine {sup 228}Ac chemical yield via ICP-MS, but {sup 133}Ba can also be used instead if ICP-MS assay is not available. Unlike some older methods, no lead or strontium holdback carriers or continual readjustment of sample pH is required.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2023

An examination of the metal ion content in the active sites of human endonucleases CPSF73 and INTS11

Human cleavage and polyadenylation specificity factor (CPSF)73 (also known as CPSF3) is the endoribonuclease that catalyzes the cleavage reaction for the 3'-end processing of pre-mRNAs. The active site of CPSF73 is located at the interface between a metallo-β-lactamase domain and a β-CASP domain. Two metal ions are coordinated by conserved residues, five His and two Asp, in the active site, and they are critical for the nuclease reaction. The metal ions have long been thought to be zinc ions, but their exact identity has not been examined. Here we present evidence from inductively coupled plasma mass spectrometry and X-ray diffraction analyses that a mixture of metal ions, including Fe, Zn, and Mn, is present in the active site of CPSF73. The abundance of the various metal ions is different in samples prepared from different expression hosts. Zinc is present at less than 20% abundance in a sample expressed in insect cells, but the sample is active in cleaving a pre-mRNA substrate in a reconstituted canonical 3'-end processing machinery. Zinc is present at 75% abundance in a sample expressed in human cells, which has comparable endonuclease activity. We also observe a mixture of metal ions in the active site of the CPSF73 homolog INTS11, the endonuclease for Integrator. Taken together, our results provide further insights into the role of metal ions in the activity of CPSF73 and INTS11 for RNA 3'-end processing.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2014

Preparation, photoluminescent properties and luminescent dynamics of BaAlF{sub 5}:Eu{sup 2+} nanophosphors

Graphical abstract: Rice-shaped BaAlF{sub 5}:Eu{sup 2+} nanophosphors were synthesized via one-pot hydrothermal process. The as-prepared BaAlF{sub 5}:Eu{sup 2+} are composed of many particles with an average diameter of 40 nm. When excited at 260 nm, the sharp line emission located at 361 nm of Eu{sup 2+} was observed. The optimum doping concentration of Eu{sup 2+} was confirmed to be 5 mol%. The strong ultraviolet emission of Eu{sup 2+} ions in BaAlF{sub 5}:Eu{sup 2+} nanoparticles suggests that these nanoparticles may have potential applications for sensing, solid-state lasers and spectrometer calibration. - Highlights: • BaAlF{sub 5}:Eu{sup 2+} nanophosphors were synthesized via a mild hydrothermal process. • The Van and Huang models were used to research the mechanism of concentration quenching. • The optimum doping concentration of Eu2+ was confirmed to be 5 mol%. - Abstract: Eu{sup 2+}-doped BaAlF{sub 5} nanophosphors were synthesized via a facile one-pot hydrothermal method. The final products were characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), and photoluminescence (PL) spectroscopy. XRD results showed that the prepared samples are single-phase. The FE-SEM and TEM images indicated that the prepared BaAlF{sub 5}:Eu{sup 2+} nanophosphors are composed of many rice-shaped particles with an average diameter of 40 nm. When excited at 260 nm, BaAlF{sub 5}:Eu{sup 2+} nanophosphors exhibit the sharp line emissions of Eu{sup 2+} at room temperature. The optimum doping concentration of Eu{sup 2+} was confirmed to be 5 mol%. The Van and Huang models were used to study the mechanism of concentration quenching and the electric dipole–dipole interaction between Eu{sup 2+} can be deduced to be a dominant for quenching fluorescence in BaAlF{sub 5}:Eu{sup 2+} nanophosphors. The strong ultraviolet emission of Eu{sup 2+} in BaAlF{sub 5}:Eu{sup 2+} nanophosphors suggests that these nanoparticles may have potential applications for sensing, spectrometer calibration and solid-state lasers.

36 MATERIALS SCIENCE↗

DOE OSTI2025

Sample glue layer investigation and mitigation for laser induced prompt impulse experiments

Understanding longer timescale material reactions under dynamic stress loading is critical for applications in materials engineering, shock physics, and planetary science. Prompt impulse experiments generate lower pressures since the ablator—the material first removed by the laser—is thicker and farther from the diagnostic plane, capturing aggregate material responses from the initial shock wave, rarefaction waves, and later time effects. This complexity demands thorough material characterization and simulation support. Since traditional sample construction is specific to supported shock experiments, designing prompt impulse experiments requires reconsideration around target design and sample engineering. Here, we present sample preparation techniques, experimental investigations, and theoretical simulations to investigate glue layer impacts, aiming to standardize samples for consistent data at lower laser fluences. We find that glue layers <30 μm have a minimal impact on peak velocity and pulse shape. The peak velocity scales linearly with glue layer thickness until a glue layer of 75 μm. For glue layers >75 μm, the peak velocity no longer scales with thickness; however, the pulse shape continues to degrade as described by simulations.

Lasers↗

DOE OSTI2024

Operando Freezing Cryogenic Electron Microscopy of Active Battery Materials

Abstract Understanding structural and chemical evolution of battery materials during operation is critical to achieving safe, efficient, and long-lasting energy storage. Cryogenic electron microscopy (cryo-EM) has become a valuable tool in battery characterization, leveraging low temperatures to improve stability of sensitive materials under electron beam irradiation. However, typical cryo-EM sample preparations leave extended time between the electrochemical point of interest and ex situ freezing of samples, during which active structures may relax, degrade, or otherwise evolve. Here, we detail a method for operando freezing cryo-EM to preserve and characterize native electrode and interfacial structures that arise during battery cycling, based on an operando plunge freezer and cold sample removal process. We validate the method on multiple electrode materials and quantify and discuss the freezing rate achieved. Operando freezing cryo-EM can be used to directly visualize transient features that arise at active electrochemical interfaces, to enable deeper understanding of structural evolution and interfacial chemistry in batteries and other electrochemical systems.

25 ENERGY STORAGE↗

DOE OSTI2019

Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils

Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ~1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S Plus Library Kit. While ssDNA viruses were identified in all three soil types, none were identified in the samples that used bead-beating, suggesting this lysis method may impact recovery. Further, 13 ssDNA vOTUs were identified compared to 582 dsDNA vOTUs, and the ssDNA vOTUs only accounted for ~4% of the assembled reads, implying dsDNA viruses were dominant in these samples. This optimized approach was combined with the previously published viral resuspension protocol into a sample-to-virome protocol for soils now available at protocols.io, where community feedback creates ‘living’ protocols. This collective approach will be particularly valuable given the high physicochemical variability of soils, which will may require considerable soil type-specific optimization. This optimized protocol provides a starting place for developing quantitatively-amplified viromic datasets and will help enable viral ecogenomic studies on organic-rich soils.

Trubl, Gareth↗

DOE OSTI2019

Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils

Trubl, Gareth↗

DOE OSTI2019

Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils

Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ∼1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S Plus Library Kit. While ssDNA viruses were identified in all three soil types, none were identified in the samples that used bead-beating, suggesting this lysis method may impact recovery. Further, 13 ssDNA vOTUs were identified compared to 582 dsDNA vOTUs, and the ssDNA vOTUs only accounted for ∼4% of the assembled reads, implying dsDNA viruses were dominant in these samples. This optimized approach was combined with the previously published viral resuspension protocol into a sample-to-virome protocol for soils now available at protocols.io , where community feedback creates ‘living’ protocols. This collective approach will be particularly valuable given the high physicochemical variability of soils, which will may require considerable soil type-specific optimization. This optimized protocol provides a starting place for developing quantitatively-amplified viromic datasets and will help enable viral ecogenomic studies on organic-rich soils.

dsDNA viruses↗