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Comparative Analysis of Two-Wheeled Mobility Options and Suitability for the U.S. Park Police

This report is a framework to help the United States Park Police in selecting and deploying various e-mobility options that are currently available in the market. In the process of creating this framework, we developed the Mobility Optimization and Deployment Engine (MODE) tool whose use could go beyond the scope of this work and mobility options.

33 ADVANCED PROPULSION SYSTEMS↗

Highest fusion performance without harmful edge energy bursts in tokamak

The path of tokamak fusion and International thermonuclear experimental reactor (ITER) is maintaining high-performance plasma to produce sufficient fusion power. This effort is hindered by the transient energy burst arising from the instabilities at the boundary of plasmas. Conventional 3D magnetic perturbations used to suppress these instabilities often degrade fusion performance and increase the risk of other instabilities. This study presents an innovative 3D field optimization approach that leverages machine learning and real-time adaptability to overcome these challenges. Implemented in the DIII-D and KSTAR tokamaks, this method has consistently achieved reactor-relevant core confinement and the highest fusion performance without triggering damaging bursts. This is enabled by advances in the physics understanding of self-organized transport in the plasma edge and machine learning techniques to optimize the 3D field spectrum. The success of automated, real-time adaptive control of such complex systems paves the way for maximizing fusion efficiency in ITER and beyond while minimizing damage to device components.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Laser Powder Bed Fusion Additive Manufacturing In-Process Monitoring and Optimization Using Thermionic Emission Detection

In this project, the emission of electrons during laser powder bed fusion additive manufacturing was identified and investigated to resolve the underlying dynamics driven by laser-material interactions. Laser powder bed fusion involves the selective laser melting of metal powder particles layer by layer which fuse together forming a larger 3D structure. Laser-based additive manufacturing approaches such as laser powder bed fusion hold the potential to revolutionize manufacturing of complex metal components in the aerospace, medical, and automotive industries. However, the difficulty in producing defect-free components by metal 3D printing is a major hurdle for widespread commercial adoption. This project resolved important dynamics in laser powder bed fusion including the identification of plasma formation mechanisms, transitions between laser-induced melting modes and stochastic precursors to defect formation. These findings were realized through implementation of electronic sensing diagnostics into an existing testbed system and harnessing capabilities developed for the time dependent analysis of laser powder bed fusion datasets. Importantly, the results advance our understanding of laser-material interactions and reveal that detection of thermionic emission can also resolve information critical to optimization of the laser powder bed fusion fabrication process. The findings are critical for advancing our fundamental understanding of the laser powder bed fusion process and can help assure the requisite fidelity of fabricated components.

36 MATERIALS SCIENCE↗

TU-EF-207-01: Introductory Remarks on Recent Advances in Breast Imaging

Breast imaging technology is advancing on several fronts. In digital mammography, the major technological trend has been on optimization of approaches for performing combined mammography and tomosynthesis using the same system. In parallel, photon-counting slot-scan mammography is now in clinical use and more efforts are directed towards further development of this approach for spectral imaging. Spectral imaging refers to simultaneous acquisition of two or more energy-windowed images. Depending on the detector and associated electronics, there are a number of ways this can be accomplished. Spectral mammography using photon-counting detectors can suppress electronic noise and importantly, it enables decomposition of the image into various material compositions of interest facilitating quantitative imaging. Spectral imaging can be particularly important in intravenously injected contrast mammography and eventually tomosynthesis. The various approaches and applications of spectral mammography are discussed. Digital breast tomosynthesis relies on the mechanical movement of the x-ray tube to acquire a number of projections in a predefined arc, typically from 9 to 25 projections over a scan angle of +/−7.5 to 25 degrees depending on the particular system. The mechanical x-ray tube motion requires relatively long acquisition time, typically between 3.7 to 25 seconds depending on the system. Moreover, mechanical scanning may have an effect on the spatial resolution due to internal x-ray filament or external mechanical vibrations. New x-ray source arrays have been developed and they are aimed at replacing the scanned x-ray tube for improved acquisition time and potentially for higher spatial resolution. The potential advantages and challenges of this approach are described. Combination of digital mammography and tomosynthesis in a single system places increased demands on certain functional aspects of the detector and overall performance, particularly in the tomosynthesis mode due to lower photon fluence per projection. This may require fast-frame acquisition and symmetric or asymmetric pixel binning in some systems. Recent studies investigated the performance of increased conversion layer thickness for contrast-enhanced imaging of the breast in dual-energy acquisition mode. In other direct conversion detectors operating in the avalanche mode, sensitivities close to the single photon response are also explored for mammography and breast tomosynthesis. The potential advantages and challenges of this approach are described. Dedicated breast CT brings x-ray imaging of the breast to true tomographic 3D imaging. It can eliminate the tissue superposition problem and does not require physical compression of the breast. Using cone beam geometry and a flat-panel detector, several hundred projections are acquired and reconstructed to near isotropic voxels. Multiplanar reconstruction facilitates viewing the breast volume in any desired orientation. Ongoing clinical studies, the current state-of-the art, and research to advance the technology are described. Learning Objectives: To understand the ongoing developments in x-ray imaging of the breast To understand the approaches and applications of spectral mammography To understand the potential advantages of distributed x-ray source arrays for digital breast tomosynthesis To understand the ongoing developments in detector technology for digital mammography and breast tomosynthesis To understand the current state-of-the-art for dedicated cone-beam breast CT and research to advance the technology. Research collaboration with Koning Corporation.

60 APPLIED LIFE SCIENCES↗

Advanced Control Methods for Particle Accelerators (ACM4PA) 2019

Los Alamos is currently developing novel particle accelerator controls and diagnostics algorithms to enable higher quality beams with lower beam losses than is currently possible. The purpose of this workshop was to consider tuning and optimization challenges of a wide range of particle accelerators including linear proton accelerators such as the Los Alamos Neutron Science Center (LANSCE), rings such as the Advanced Photon Source (APS) synchrotron, free electron lasers (FEL) such as the Linac Coherent Light Source (LCLS) and LCLS-II, the European X-ray Free Electron Laser (EuXFEL), the Swiss FEL, and the planned MaRIE FEL, and plasma wake- eld accelerators such as FACET, FACET-II, and AWAKE at CERN. One major challenge is an the ability to quickly create very high quality, extremely intense, custom current and energy pro le beams while working with limited real time non-invasive diagnostics and utilizing time-varying uncertain initial beam distributions and accelerator components. Currently, a few individual accelerator labs have been developing and applying their own diagnostics tools and custom control and ML algorithms for au- tomated machine tuning and optimization. The goal of this workshop was to bring together a group of accelerator physicists and accelerator related control and ML experts in order to de ne which controls and diagnostics would be most useful for existing and future accelerators and to create a plan for developing a new family of algorithms that can be shared and maintained by the community.

43 PARTICLE ACCELERATORS↗

Review of SiC material development for nuclear fusion applications: Cross-cutting research and emerging opportunities

The SiC-based materials, particularly SiC-fiber-reinforced SiC matrix (SiC/SiC) composites, show strong potential for structural and functional applications in future fusion power plants because they can operate at high temperatures with a range of coolants and breeders, thereby enabling higher energy conversion efficiency. Here, this paper presents recent advancements in the development of SiC-based materials, focusing on processing techniques and material performance and resistance under fusion-relevant environments. The processing activities have emphasized near-net-shape fabrication and the joining of SiC subcomponents, with processing methods and material compositions informed by previous irradiation experiments on various grades of SiC. Research on irradiation effects has remained focused on degradation mechanisms and the microstructural optimization of SiC/SiC composites irradiated to high neutron damage levels. Analysis of irradiation defects in SiC has advanced via the application of cutting-edge characterization methods, among which Raman spectroscopy is becoming a common tool to assess atomic-scale chemical disorder. Fusion–fission crosscutting irradiation research has explored combined effects in SiC/SiC composites with application-relevant geometries, including bowing of SiC/SiC composite channels under neutron flux gradients, stress evolution in SiC/SiC composite tubes under through-thickness temperature gradients, and irradiation-enhanced corrosion in SiC. Finally, research opportunities for component testing and assessment under fusion-relevant conditions, in support of emerging concepts from the private fusion sector, are discussed.

Advanced manufacturing↗

Computationally Efficient Control Co-Design Optimization Framework with Mixed-Fidelity Fluid and Structure Analysis: OpenTurbineCoDe (OTCD) - ARPA-E ATLANTIS Phase I Project (Final Technical Report)

The goal of the Department of Energy (DOE) Advanced Research Projects Agency-Energy (ARPA-E) Aerodynamic Turbines Lighter and Afloat with Nautical Technologies and Integrated Servo-control (ATLANTIS) Program is to develop new technologies for floating offshore wind turbines, or wind farms, using the discipline of control co-design (CCD.) In this context, our goal is to develop a computationally efficient optimization framework for design of floating offshore wind turbines. Our specific aim is to utilize high-fidelity structural, aerodynamic, aero-structural tools, and to derive control-oriented reduced- or low-order models directly from the high-fidelity tools. We proposed a mixed-fidelity modeling approach which means that we are also using low- and mid-fidelity tools when necessary. This research is conducted by a multidisciplinary team consisting of Rutgers University, University of Michigan, Brigham Young University, and the National Renewable Energy Laboratory (NREL). The computational framework, called OpenTurbineCoDe, is designed to integrate, where possible, traditional structural, aerodynamic, aeroelastic models (e.g., OpenFAST) and advanced control algorithms with higher fidelity simulation tools including Reynolds-averaged Navier–Stokes (RANS) solvers, and three-dimensional structural finite element solvers. All the high-fidelity tools used in this research provide numerically exact gradients to facilitate both efficient optimization and local linearization for control implementation.

17 WIND ENERGY↗

Perspectives on Polyolefin Catalysis in Microfluidics for High-Throughput Screening: A Minireview

Polyolefins are the largest produced plastics in the world which traditionally employ continuous stirred tank reactors and fluidized bed reactors for commercial production. The operating condition, reaction kinetics, and molecular interactions inside the reactor strongly affect the polyolefin properties, which require stringent process control in conventional procedures. Understanding the catalytic pathway, behavior of polymer particles and effect of reactor conditions are essential for designing specific polymer properties, namely the molecular weight, chain length, polydispersity, etc. Microfluidics can play a significant role in designing polymers tailored to the user needs. Smaller channel dimensions help obtain uniform reaction conditions over the length of the microfluidic reactor in a controlled environment. With real-time monitoring techniques in microfluidics, even single particle growth of polymer can be studied to understand the parameters affecting the polymer properties. High throughput microfluidics can help catalyst screening in a short duration with less consumption of reagents generating less waste. When supplemented with efficient machine learning algorithms, automated high throughput microfluidics has the potential to rapidly optimize the process and develop new knowledge even with a limited data set. When trained on data sets generated using microfluidic experiments that are designed efficiently with working knowledge of the process, machine learning algorithms can provide the relationship between the multivariable parameters space and polymer properties, which is not possible with the traditional statistical methods and interpolation techniques. Here, the rise in the utilization of microfluidics, with the advancement of machine learning algorithms, for polyolefin catalysis, highlights the importance of microfluidics for catalyst discovery, parameter optimization, and understanding reaction pathway for producing polymers with specific properties for specialized applications.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Structural Loads Analysis for Wave Energy Converters: Preprint

This study explores and verifies the generalized body-modes method for evaluating the structural loads on a wave energy converter (WEC). Historically, WEC design methodologies have focused primarily on accurately evaluating hydrodynamic loads, while methodologies for evaluating structural loads have yet to be fully considered and incorporated into the WEC design process. As wave energy technologies continue to advance, however, it has become increasingly evident that an accurate evaluation of the structural loads will enable an optimized structural design, as well as the potential utilization of composites and flexible materials, and hence reduce WEC costs. Although there are many computational fluid dynamics, structural analyses and fluid-structure-interaction (FSI) codes available, the application of these codes is typically too computationally intensive to be practical in the early stages of the WEC design process. The generalized body-modes method, however, is a reduced order, linearized, frequency-domain FSI approach, performed in conjunction with the linear hydrodynamic analysis, with computation times that could realistically be incorporated into the WEC design process. The objective of this study is to verify the generalized body-modes approach in comparison to high-fidelity FSI simulations to accurately predict structural deflections and stress loads in a WEC. Two verification cases are considered, a free-floating barge and a fixed-bottom column. Details for both the generalized body-modes models and FSI models are first provided. Results for each of the models are then compared and discussed. Finally, based on the verification results obtained, future plans for incorporating the generalized body-modes method into the WEC simulation tool, WEC-Sim, and the overall WEC design process are discussed.

17 WIND ENERGY↗

Coupling Big Data Analytics and Reactive Transport Modeling for Cost-effective Groundwater Monitoring - 17163

This study presents an innovative approach for sustainable and cost-effective groundwater monitoring. This approach takes advantage of recent advances in various technologies: (1) in situ autonomous sensors, (2) big data analytics, and (3) parallel high-performance computing for flow and reactive transport modeling. In situ sensors are used to periodically measure the key variables (such as pH, redox potential, electrical conductivity, and groundwater level), which control contaminant mobility and the plume spatial and temporal distribution. Based on a limited number of groundwater sampling, the data analytics methods-data mining and machine learning-allow us to identify and quantify the correlations between the in situ-measured variables and contaminant concentrations, and also to detect significant changes associated with the plume mobility. In addition, a state-of-the-art parallel numerical flow and reactive transport simulator Amanzi, and uncertainty quantification software Agni are used to provide an improved physical and mechanistic understanding of the contaminated groundwater system behavior, and to predict the long-term plume distribution for optimizing and adapting the monitoring strategy. Amanzi and Agni were developed as part of the Advanced Simulation Capability for Environmental Management (ASCEM) program of the DOE Office of Environmental Management. Such modeling is critical, particularly, for assessing the impact of climate change and associated hydrological shifts. The developed approach is expected to significantly reduce the groundwater sampling frequency and associated cost. In addition, the real-time information on plume mobility serves as an early warning system, improving the resiliency of contaminated or potentially contaminated sites. We demonstrate this approach using as an example the Savannah River Site (SRS) F-Area, where groundwater is contaminated by various radionuclides, including uranium, tritium and technetium. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Characterization of Large Batch 14YWT Heat (NFA-2) for JOYO Irradiations (Rev.1)

The development and deployment of advanced cladding materials in advanced reactors requires the collection of relevant performance data for each cladding concept. Novel cladding concepts typically undergo significant innovation and iteration over the course of their development, and intention should be given to providing a snapshot of present performance. Thus, a modern heat of the nanostructured ferritic alloy (NFA) 14YWT is produced using state-of-the-art procedures for impurity minimization combined with thermomechanical processing methods designed to optimize fracture toughness, tensile strength, and ductility. This new heat of 14YWT, NFA-2, will provide the Advanced Fuels Campaign with adequate material for upcoming irradiation and mechanical testing campaigns. This report outlines procurement of new gas atomized 14YWT powder, production of four new bars of 14YWT, and subsequent mechanical and microstructural property characterization.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

AVESTAR Center for Operational Excellence of Electricity Generation Plants

To address industry challenges in attaining operational excellence for electricity generation plants, the U.S. Department of Energy’s (DOE) National Energy Technology Laboratory (NETL) has launched a world-class facility for Advanced Virtual Energy Simulation Training and Research (AVESTARTM). This presentation will highlight the AVESTARTM Center simulators, facilities, and comprehensive training, education, and research programs focused on the operation and control of high-efficiency, near-zero-emission electricity generation plants. The AVESTAR Center brings together state-of-the-art, real-time, high-fidelity dynamic simulators with full-scope operator training systems (OTSs) and 3D virtual immersive training systems (ITSs) into an integrated energy plant and control room environment. AVESTAR’s initial offering combines--for the first time--a “gasification with CO2 capture” process simulator with a “combined-cycle” power simulator together in a single OTS/ITS solution for an integrated gasification combined cycle (IGCC) power plant with carbon dioxide (CO2) capture. IGCC systems are an attractive technology option for power generation, especially when capturing and storing CO2 is necessary to satisfy emission targets. The AVESTAR training program offers a variety of courses that merge classroom learning, simulator-based OTS learning in a control-room operations environment, and immersive learning in the interactive 3D virtual plant environment or ITS. All of the courses introduce trainees to base-load plant operation, control, startups, and shutdowns. Advanced courses require participants to become familiar with coordinated control, fuel switching, power-demand load shedding, and load following, as well as to problem solve equipment and process malfunctions. Designed to ensure work force development, training is offered for control room and plant field operators, as well as engineers and managers. Such comprehensive simulator-based instruction allows for realistic training without compromising worker, equipment, and environmental safety. It also better prepares operators and engineers to manage the plant closer to economic constraints while minimizing or avoiding the impact of any potentially harmful, wasteful, or inefficient events. The AVESTAR Center is also used to augment graduate and undergraduate engineering education in the areas of process simulation, dynamics, control, and safety. Students and researchers gain hands-on simulator-based training experience and learn how the commercial-scale power plants respond dynamically to changes in manipulated inputs, such as coal feed flow rate and power demand. Students also analyze how the regulatory control system impacts power plant performance and stability. In addition, students practice start-up, shutdown, and malfunction scenarios. The 3D virtual ITSs are used for plant familiarization, walk-through, equipment animations, and safety scenarios. To further leverage the AVESTAR facilities and simulators, NETL and its university partners are pursuing an innovative and collaborative R&D program. In the area of process control, AVESTAR researchers are developing enhanced strategies for regulatory control and coordinated plant-wide control, including gasifier and gas turbine lead, as well as advanced process control using model predictive control (MPC) techniques. Other AVESTAR R&D focus areas include high-fidelity equipment modeling using partial differential equations, dynamic reduced order modeling, optimal sensor placement, 3D virtual plant simulation, and modern grid. NETL and its partners plan to continue building the AVESTAR portfolio of dynamic simulators, immersive training systems, and advanced research capabilities to satisfy industry’s growing need for training and experience with the operation and control of clean energy plants. Future dynamic simulators under development include natural gas combined cycle (NGCC) and supercritical pulverized coal (SCPC) plants with post-combustion CO2 capture. These dynamic simulators are targeted for use in establishing a Virtual Carbon Capture Center (VCCC), similar in concept to the DOE’s National Carbon Capture Center for slipstream testing. The VCCC will enable developers of CO2 capture technologies to integrate, test, and optimize the operation of their dynamic capture models within the context of baseline power plant dynamic models. The objective is to provide hands-on, simulator-based “learn-by-operating” test platforms to accelerate the scale-up and deployment of CO2 capture technologies. Future AVESTAR plans also include pursuing R&D on the dynamics, operation, and control of integrated electricity generation and storage systems for the modern grid era. Special emphasis will be given to combining load-following energy plants with renewable and distributed generating supplies and fast-ramping energy storage systems to provide near constant baseload power.

97 MATHEMATICS AND COMPUTING↗

Extracting and Generating PV Soiling Profiles for Analysis, Forecasting, and Cleaning Optimization

The identification and prediction of the daily soiling profiles of a photovoltaic site is essential to plan the optimal cleaning schedule. In this article, we analyze and propose various methods to extract and generate photovoltaic soiling profiles, in order to improve the analysis and the forecast of the losses. New soiling rate extraction methods are proposed to reflect the seasonal variability of the soiling rates and, for this reason, are found to identify the most convenient cleaning day with the highest accuracy for the investigated sites. Also, we present an approach that could be used to predict future soiling losses through the implementation of stochastic weather generation algorithms whose ability to identify in advance the best cleaning schedule is also successfully tested. The methods presented in this article can optimize the operation and maintenance schedule and could make it possible, in the future, to predict soiling losses through analysis based only on environmental parameters, such as rainfall and particulate matter, without the need of long-term soiling data.

14 SOLAR ENERGY↗

Risk-Informed Safety Margins Characterization (RISMC) Pathway Technical Program Plan

Safety is central to the design, licensing, operation, and economics of nuclear power plants (NPPs). As the current Light Water Reactor (LWR) NPPs age beyond 60 years, there are possibilities for increased frequency of systems, structures, and components (SSCs) degradations or failures that initiate safety-significant events, reduce existing accident mitigation capabilities, or create new failure modes. Plant designers commonly “over-design” portions of NPPs and provide robustness in the form of redundant and diverse engineered safety features to ensure that, even in the case of well-beyond design basis scenarios, public health and safety will be protected with a very high degree of assurance. This form of defense-in-depth is a reasoned response to uncertainties and is often referred to generically as “safety margin.” Historically, specific safety margin provisions have been formulated, primarily based on “engineering judgment.” The ability to better characterize (i.e., describe and quantify) safety margin is important to improved decision making about LWR design and long-term operation. A systematic approach to characterizing safety margins and the subsequent risk informed margins management options represents a vital input to the licensee and regulatory analysis and decision making that will be involved. In addition, as research and development (R&D) in the LWRS Program and other collaborative efforts yield new data and improved scientific understanding of physical processes that govern the aging and degradation of plant SSCs (and concurrently support technological advances in nuclear reactor fuels and plant instrumentation and control systems), needs and opportunities to better optimize plant safety, economics, and performance will become known. The purpose of the Risk-Informed Safety Margins Characterization (RISMC) Pathway R&D is to support plant decisions for risk-informed margins management with the aim to improve economics, reliability, and sustain safety of current NPPs over periods of extended plant operations. The goals of the RISMC Pathway are twofold: (1) develop and demonstrate a risk-assessment method that is coupled to safety margin quantification that can be used by NPP decision makers as part of risk-informed margin management strategies; (2) create an advanced RISMC Toolkit that enables more accurate representation of NPP safety margins. The methods and tools provided by RISMC are essential to a comprehensive and integrated risk-informed margin management approach that supports effective preservation of margin for both active and passive SSCs.

97 MATHEMATICS AND COMPUTING↗

A Comprehensive Comparison of Methods for Evaluating Dispatch of Long-Duration Energy Storage in Power Systems Models

Long-duration energy storage (LDES) could play a pivotal role in the transformation of electricity grids with high shares of variable renewable energy (VRE) such as solar and wind. However, the weather-dependent nature of VRE introduces challenges for grid balancing and stability, which LDES - along with short-duration energy storage (SDES) - can help address. However, modeling LDES in production cost models (PCMs) is particularly challenging due to the need for high temporal resolution over extended optimization windows while preserving chronology, which ensures the alignment of energy storage operations with VRE generation over multi-day periods. This report compares traditional dispatch methods with advanced LDES dispatch strategies, such as the extended horizon approach, across different PCM platforms and examines tradeoffs and scalability. The comparison reveals that the traditional 1-day optimization horizon within the PCM leads to inefficient utilization of LDES. In contrast, extending the optimization horizon as much as possible significantly reduces curtailment and improves storage dispatch, especially in renewable-dense systems. There is also promise in using state-of-charge or end volume targets set by an external model, however this requires an additional modeling set and generally increases computational burden. This paper presents a comparison of these various methods in a number of power systems, showing algorithms initially in small test systems and scaling up to large, country-wide simulations. Overall, the research presents the trade-offs of various computational methods and illustrates how LDES may play an essential role in power systems of the future.

14 SOLAR ENERGY↗

Optimizing de novo genome assembly from PCR-amplified metagenomes

Background. Metagenomics has transformed our understanding of microbial diversity across ecosystems, with recent advances enabling de novo assembly of genomes from metagenomes. These metagenome-assembled genomes are critical to provide ecological, evolutionary, and metabolic context for all the microbes and viruses yet to be cultivated. Metagenomes can now be generated from nanogram to subnanogram amounts of DNA. However, these libraries require several rounds of PCR amplification before sequencing, and recent data suggest these typically yield smaller and more fragmented assemblies than regular metagenomes. Methods. Here we evaluate de novo assembly methods of 169 PCR-amplified metagenomes, including 25 for which an unamplified counterpart is available, to optimize specific assembly approaches for PCR-amplified libraries. We first evaluated coverage bias by mapping reads from PCR-amplified metagenomes onto reference contigs obtained from unamplified metagenomes of the same samples. Then, we compared different assembly pipelines in terms of assembly size (number of bp in contigs ≥ 10kb) and error rates to evaluate which are the best suited for PCR-amplified metagenomes. Results. Read mapping analyses revealed that the depth of coverage within individual genomes is significantly more uneven in PCR-amplified datasets versus unamplified metagenomes, with regions of high depth of coverage enriched in short inserts. This enrichment scales with the number of PCR cycles performed, and is presumably due to preferential amplification of short inserts. Standard assembly pipelines are confounded by this type of coverage unevenness, so we evaluated other assembly options to mitigate these issues. We found that a pipeline combining read deduplication and an assembly algorithm originally designed to recover genomes from libraries generated after whole genome amplification (single-cell SPAdes) frequently improved assembly of contigs ≥ 10kb by 10 to 100-fold for low input metagenomes. Conclusions. PCR-amplified metagenomes have enabled scientists to explore communities traditionally challenging to describe, including some with extremely low biomass or from which DNA is particularly difficult to extract. Here we show that a modified assembly pipeline can lead to an improved de novo genome assembly from PCR-amplified datasets, and enables a better genome recovery from low input metagenomes.

Roux, Simon↗

Optimizing de novo genome assembly from PCR-amplified metagenomes

Background: Metagenomics has transformed our understanding of microbial diversity across ecosystems, with recent advances enabling de novo assembly of genomes from metagenomes. These metagenome-assembled genomes are critical to provide ecological, evolutionary, and metabolic context for all the microbes and viruses yet to be cultivated. Metagenomes can now be generated from nanogram to subnanogram amounts of DNA. However, these libraries require several rounds of PCR amplification before sequencing, and recent data suggest these typically yield smaller and more fragmented assemblies than regular metagenomes. Methods: Here we evaluate de novo assembly methods of 169 PCR-amplified metagenomes, including 25 for which an unamplified counterpart is available, to optimize specific assembly approaches for PCR-amplified libraries. We first evaluated coverage bias by mapping reads from PCR-amplified metagenomes onto reference contigs obtained from unamplified metagenomes of the same samples. Then, we compared different assembly pipelines in terms of assembly size (number of bp in contigs ≥ 10kb) and error rates to evaluate which are the best suited for PCR-amplified metagenomes. Results: . Read mapping analyses revealed that the depth of coverage within individual genomes is significantly more uneven in PCR-amplified datasets versus unamplified metagenomes, with regions of high depth of coverage enriched in short inserts. This enrichment scales with the number of PCR cycles performed, and is presumably due to preferential amplification of short inserts. Standard assembly pipelines are confounded by this type of coverage unevenness, so we evaluated other assembly options to mitigate these issues. We found that a pipeline combining read deduplication and an assembly algorithm originally designed to recover genomes from libraries generated after whole genome amplification (single-cell SPAdes) frequently improved assembly of contigs ≥ 10kb by 10 to 100-fold for low input metagenomes. Conclusions: PCR-amplified metagenomes have enabled scientists to explore communities traditionally challenging to describe, including some with extremely low biomass or from which DNA is particularly difficult to extract. Here we show that a modified assembly pipeline can lead to an improved de novo genome assembly from PCR-amplified datasets, and enables a better genome recovery from low input metagenomes.

Roux, Simon↗

Optimizing de novo genome assembly from PCR-amplified metagenomes

Background. Metagenomics has transformed our understanding of microbial diversity across ecosystems, with recent advances enabling de novo assembly of genomes from metagenomes. These metagenome-assembled genomes are critical to provide ecological, evolutionary, and metabolic context for all the microbes and viruses yet to be cultivated. Metagenomes can now be generated from nanogram to subnanogram amounts of DNA. However, these libraries require several rounds of PCR amplification before sequencing, and recent data suggest these typically yield smaller and more fragmented assemblies than regular metagenomes. Methods. Here we evaluate de novo assembly methods of 169 PCR-amplified metagenomes, including 25 for which an unamplified counterpart is available, to optimize specific assembly approaches for PCR-amplified libraries. We first evaluated coverage bias by mapping reads from PCR-amplified metagenomes onto reference contigs obtained from unamplified metagenomes of the same samples. Then, we compared different assembly pipelines in terms of assembly size (number of bp in contigs ≥ 10 kb) and error rates to evaluate which are the best suited for PCR-amplified metagenomes. Results. Read mapping analyses revealed that the depth of coverage within individual genomes is significantly more uneven in PCR-amplified datasets versus unamplified metagenomes, with regions of high depth of coverage enriched in short inserts. This enrichment scales with the number of PCR cycles performed, and is presumably due to preferential amplification of short inserts. Standard assembly pipelines are confounded by this type of coverage unevenness, so we evaluated other assembly options to mitigate these issues. We found that a pipeline combining read deduplication and an assembly algorithm originally designed to recover genomes from libraries generated after whole genome amplification (single-cell SPAdes) frequently improved assembly of contigs ≥10 kb by 10 to 100-fold for low input metagenomes. Conclusions. PCR-amplified metagenomes have enabled scientists to explore communities traditionally challenging to describe, including some with extremely low biomass or from which DNA is particularly difficult to extract. Here we show that a modified assembly pipeline can lead to an improved de novo genome assembly from PCR-amplified datasets, and enables a better genome recovery from low input metagenomes.

59 BASIC BIOLOGICAL SCIENCES↗