Harcourt J, Tamin A, Lu X, Kamili S, Sakthivel SK, Murray J, et al. All other genomes were obtained from NCBI. TapeStation Software for NGS Sample Quality Control | Agilent conceived and designed the experiments, conducted experiments, analyzed data, and wrote the manuscript; K.B.B. S2, Supplemental Tables14). A number of different approaches have been used to sequence SARS-CoV-2. It is suitable to analyze size, quantity, and integrity of your samples. 7(2), 1118 (2010). The Agilent TapeStation system is an automated electrophoresis solution for the sample quality control of DNA and RNA samples. 3a for 25 or 35 PCR cycles using tailed versions of the ARTIC v3 primers split into two separate pools. BMC Genomics Supplemental Table2. Supplemental Fig. Comparison of sequence capture, ARTIC v3 amplicon, and tailed amplicon workflows on clinical specimens spanning a range of viral loads. Nature. New! Any one have suggestions for alternative systems for analyzing fragment sizes (other than gels)? There was complete concordance in the variant calls for all samples with N1 and N2 Ct values below 30, but less agreement among variant calls between methods for the sample with N1 and N2 Ct values of approximately 35 (Fig. Bioinformatics. 105(8), 10439 (2015). In the future, it will be interesting to determine the absolute sequencing limit of this method. Researchers have used enrichment strategies to increase the number of target reads in sequencing. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The primers for the primary amplification contained both SARS-CoV-2 targeting sequences (derived from the ARTIC v3 designs), as well as adapter tails for adding indices and Illumina flow cell adapters in a secondary amplification. 2). The library preparations were performed according to the SureSelect XT HS Target Enrichment System for Illumina Paired-End Multiplexed Sequencing Library protocol (Version A1, July 2017). However, the relatively low-cost of amplicon methods make them a good choice for population-scale viral surveillance and such approaches have recently been used successfully to monitor the spread of viruses such as Zika and Ebola [2,3,4]. Supplemental Fig. My Agilent Bioanalyzer is giving me fits lately! Whole genome sequencing approaches will provide more precise molecular characterization of the diversity among populations. Over the past ten years, NGS (next generation sequencing) has been widely applied to identity pathogens, characterize genetic variants, and provide a molecular basis for building additional diagnostic tools. PubMedGoogle Scholar. The RNA ScreenTape system is designed for analyzing eukaryote and Physical Specifications Signal- to- noise >3 (single peak) Measured against 2200 TapeStation System Agilent Technologies Storage Conditions The PCR products from pool 1 and pool 2 for each sample were combined and then diluted 1:100 in sterile, nuclease-free water, and a second PCR reaction was set up to add the Illumina flow cell adapters and indices. For me the Experion system was more forgiving when it came to chip loading. Hashino M, Tanaka R, Kuroda M. A proposal of alternative primers for the ARTIC Network's multiplex PCR to improve coverage of SARS-CoV . 2015;523:21720. Issue: When using the Agilent 4200, 4150 and 2200 TapeStation systems, the DV 200 of FFPE RNA samples can be calculated within the TapeStation analysis software. 3c, Supplemental Fig. 30(14), 20682069 (2014). We thank California Department of Food and Agriculture (CDFA) for providing the infected citrus samples. 2a-b, Supplemental Tables14). The hybridized libraries were purified with Dynabeads MyOne Streptavidin T1 magnetic beads (ThermoFisher Scientific, Waltham, MA), then the beads with captured DNA were washed one time with wash buffer 1 and five times with wash buffer 2 to remove non-specific binding. Here we describe a low-cost, streamlined, all amplicon-based method for sequencing SARS-CoV-2, which bypasses costly and time-consuming library preparation steps. A rapid, cost-effective tailed amplicon method for sequencing SARS-CoV-2, https://doi.org/10.1186/s12864-020-07283-6, https://www.protocols.io/view/sars-cov-2-tailed-amplicon-illumina-sequencing-bipikdke, https://doi.org/10.1186/s13059-018-1618-7, https://doi.org/10.1038/s41579-020-0354-7, https://doi.org/10.1093/bioinformatics/bty407, https://doi.org/10.1016/j.cub.2020.03.022, https://doi.org/10.1101/2020.08.25.265074, https://doi.org/10.1101/2020.03.10.985150, https://doi.org/10.1186/s13059-019-1691-6, http://www.bioinformatics.babraham.ac.uk/projects/fastqc/, https://doi.org/10.1093/bioinformatics/btt593, https://doi.org/10.1093/bioinformatics/btp698, http://creativecommons.org/licenses/by/4.0/, http://creativecommons.org/publicdomain/zero/1.0/. The poorer performance with respect to coverage metrics with the tailed amplicon v1 protocol was due to substantially worse balance between the different tiled amplicons compared with the ARTIC v3 (untailed) primers (Fig. Arrow indicates primer dimers on gel. 130 Biotechnology Building Nat Biotechnol. Improved high-molecular-weight DNA extraction, nanopore sequencing and A-F) Observed read depth for each of the expected amplicons for the indicated sample amplified with the ARTIC v3 protocol with TruSeq library preparation at a subsampled read depth of 100,000 raw reads. TapeStation instruments for DNA & RNA Quality Control | Agilent Correspondence to Tailed amplicon v1 pool primer sequences. The IRB panel used WORKSHEET: Human Research (HRP-310) to make the determination that this study was exempt as not human research as defined by DHHS regulations. Thorvaldsdttir, H., Robinson, J. T. & Mesirov, J. P. Integrative Genomics Viewer (IGV): high-performance genomics data visualization and exploration. This Agilent tape station can scale easily between tube strips, and 96 well plate formats, capturing results within one minute per sample. Characterization of Candidatus Liberibacter asiaticus populations by double-locus analyses. Bioanalyzer & TapeStation - Biopolymers Facility For target selection, pre-designed probes are added to the mixed genomic DNA extracts and capture their complimentary DNA sequences through complimentary hybridization, allowing the uncaptured DNA to be removed during wash steps. RNA from sample (UMGC-6) was completely consumed in initial testing and could not be compared across all methods. Comes in most handy when a customer gives us a library that is "200-400 bases-I swear" and nothing shows up on Tape Station High Sens DNA Assay. The tree with the highest likelihood across 10 runs was selected. Phylogenies were generated with all samples and 11 published genomes (TableS2) using two methods, core SNPs and the pan-genome. 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Huanglongbing (HLB), or citrus greening, is a devastating citrus disease caused by phloem-restricted gram-negative bacteria Candidatus Liberibacter spp1,2. Liberibacter asiaticus (CLas) is the most widespread and is the only species associated with the disease in the United States (U.S.)4. Ca. General. Double-stranded cDNA size was determined using Bioanalyzer high sensitivity DNA assay (Agilent, Santa Clara, CA) and quantified with Qubit Fluorometer and High Sensitivity DNA assay (Thermo Fisher Scientific, Waltham, MA). Cryptic transmission of SARS-CoV-2 in Washington state. Cycling conditions were: 98C for 30s, followed by 25 or 35cycles of 98C for 15s and 65C for 5min. Without special enrichment, NGS can rarely detect low copy number pathogen sequences from complex samples due to low pathogen/host nucleic acid ratio. After SureSelect enrichment, both of these samples had 99% genome coverage with at least 250X depth of coverage. Devices from other companies that anyone can recommend? Curr Biol. Bankevich, A. et al. Effective disease managing efforts require a greater understanding of the causal agents, which can be achieved through whole genome sequencing. In general, the same regions were not always missing, with only ~2kb shared sites missing across samples. The following reaction was set up to create cDNA using the ARTIC v3 protocol: 5L template RNA, 11L nuclease-free water, 4L SuperScript IV VILO master mix (Thermo Fisher Scientific, Waltham, MA). Nine samples spanning a range of viral loads as assessed by the Ct values of the viral N1 and N2 targets by qRT-PCR were selected for these studies. Bedford T, Greninger AL, Roychoudhury P, Starita LM, Famulare M, Huang M-L, et al. The Fragment Analyzer systems utilize automated parallel capillary electrophoresis to provide reliable quality control (QC) for nucleic acids. However, NGS technology has significant limitations when performing pathogen diagnostics in complex metagenomic samples. Complete genome sequence of citrus huanglongbing bacterium, Candidatus Liberibacter asiaticus obtained through metagenomics. (Lonza's FlashGel is a similar system.) MathSciNet 308(2), 256262 (2018). 4200 TapeStation System (Agilent) - We use this instrument as an alternative to the Fragment Analyzer as part of some of our library preparation workflows. The slightly lower coverage metrics at a given subsampled read depth for the tailed amplicon v2 method can likely be explained by primer dimer formation during the two-step amplification process, which is more pronounced for higher N1 and N2 Ct samples (Supplemental Fig. Nat Med. CAS The DV 200 score is a quality score for evaluating quality of RNA derived from formalin-fixed paraffin-embedded (FFPE) samples established by Illumina Inc. in 2016. . The first CLas genome sequence was released in 2009, isolated from a single infected psyllid13, and in nearly 10 years since there have been only 14 additional CLas genomes deposited to NCBI (only five are complete). The overall workflow is depicted in Fig. Supplemental Fig. Through an iterative testing process, we demonstrate that with the tailed amplicon v2 method, a four-pool amplification scheme produces data with comparable amplicon balance, coverage metrics, and variant calls to the ARTIC v3 approach. We describe a modified workflow for SARS-CoV-2 sequencing which builds on the tiled amplicon approach developed by the ARTIC consortium and currently employed by many labs around the world. After all wash steps, the beads were suspended in 50l of nuclease free water. Wylie, T. N., Wylie, K. M., Herter, B. N. & Storch, G. A. Without enrichment, LHCA-20 and SGCA-20, the highest pathogen concentration samples, had genome coverage of 65 and 60%, respectively, both with 1x depth of coverage (Table1). cDNA synthesis reactions were incubated at: 25C for 10min, followed by 50C for 10min and 85C for 5min. Ithaca, NY 14853Email us. Genome Announc. Not surprisingly, we got the same prophage pattern for the SGCA strain sequenced in this study as SGCA5 (SC1 only), another strain from the same location14. Names of CLas samples were listed on the left. With its unique design and intuitive features, common QC bottlenecks are resolved by the automation of key steps such as gel loading and sample injection increasing lab efficiency. 1b), in which cDNA is made from SARS-CoV-2 positive samples and amplified using primers that generate tiled PCR products are being used to sequence SARS-CoV-2 [3]. 2020;26.1266-73. Genes | Free Full-Text | Evaluation of the Ion AmpliSeq SARS-CoV-2 Use the Previous and Next buttons to navigate the slides or the slide controller buttons at the end to navigate through each slide. The Agilent TapeStation is used for DNA analysis. All four LHCA samples are also clustered together. A total of 100ng of amplicons from the ARTIC protocol were used as the input for library preparation. Five patient samples with N1 and N2 Ct values ranging from ~2035 and the BEI WA isolate sample were selected for TruSeq library prep and sequencing; one sample (N1 Ct=20, N2 Ct=20.4) was prepared in triplicate. Genome Biol. Phylogenic tree (ML midpoint rooted tree) of 849 core SNVs of Candidatus Liberibacter asiaticus strains generated with Rax Maximum Likelihood method. We have the Tape Station for Agilent. Samples were processed as described above for the two-pool tailed amplicon sequencing workflow, with the exception that in the first round of PCR, four separate reactions were set up using primer pools 1.1, 1.2, 2.1, and 2.2 (see Supplemental Data File2 for primer sequences and pool composition) using 2.5L of template cDNA per reaction. While other groups in the company chose the BioA for the sake of "it's the standard," we chose the Advanced Analytical as it outperformed in almost every way, including running fragment analysis of dirty digests, without getting clogged. Phytopathology, https://doi.org/10.1094/PHYTO-06-18-0185-R (2018). Agilent 4200 TapeStation System TechWiz4u 41 subscribers Subscribe 20K views 6 years ago New Agilent 4200 TapeStation For RNA and DNA analysis. A detailed protocol is available on protocols.io: https://www.protocols.io/view/sars-cov-2-tailed-amplicon-illumina-sequencing-bipikdke. I am a little hesitant to recommend this as the machine is nearly 2 decades old now. G) 2% agarose gel showing the presence of primer dimers particularly in high N1/N2 Ct samples when indexed using different PCR cycling conditions. 2020;2019:2020.04.02.022186. In this study, we assess the ability of a target enrichment method, Agilent SureSelect XT HS (hereafter referred to as SureSelect), to enrich CLas genomic DNA from infected citrus genomic DNA, and in turn greatly reduce the cost and increase the coverage and reliability of whole genome sequencing. The ARTIC primer pools have gone through multiple iterations to improve evenness of coverage [13]. We first evaluated the different SARS-CoV-2 sequencing workflows in their performance with a previously sequenced SARS-CoV-2 isolate strain from Washington state (2019-nCoV/USA-WA1/2020) provided by BEI Resources [15]. Gohl, D.M., Garbe, J., Grady, P. et al. 22, 10111020 (2009). Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. 1). The Agilent 4200 TapeStation system (G2991AA) is an automated platform for scalable, flexible, faster and more reliable electrophoresis. 10L of PCR product for each sample was normalized using a SequalPrep 96-well Normalization Plate Kit (Thermo Fisher Scientific, Waltham, MA). 2023 BioMed Central Ltd unless otherwise stated. 2a-b, Supplemental Tables12). Interested in learning more about the TapeStation systems and how easy-to-use ScreenTape technology can give you a faster time to results and constant per-sample costs in sample quality control? The disease has since been identified in multiple states (USDA APHIS Citrus Greening Quarantine map, https://www.aphis.usda.gov/plant_health/plant_pest_info/citrus_greening/downloads/pdf_files/nationalquarantinemap.pdf). The system includes instrument, software, reagents, and ScreenTape devices to analyze size, quantity, and integrity of your DNA and RNA sample. CLas associated HLB was first found in Florida in early September, 20055 and was vectored by the Asian citrus psyllid (Diaphorina citri), which had been introduced into Florida in the late 1990s. Agilent 2200 TapeStation - Boston Laboratory Equipment Croucher, N. J. et al. The integrity of the extracted RNA was analyzed using the Agilent high sensitivity RNA screentape assay on Agilent 2200 TapeStation following the manufacturer's guidelines (Agilent, Santa Clara, CA). 8-well PCR tube strips or 96-well sample plates are available depending on sample throughput, bringing added flexibility. The system integrates an instrument, data processing software, reagents, and ScreenTape devices specific for DNA and RNA. Consistent with previous descriptions of the ARTIC v3 primers, the balance between the tiled amplicons across these samples was relatively even, with a mean CV of 0.61 among the five patient samples tested, and 0.55 for samples with a N1 and N2 Ct of less than 30 (Fig. PLoS One, https://doi.org/10.1371/journal.pone.0112968 (2014). Cai, W., Yan, Z., Rascoe, J. The two SGCA strain samples are clustered together and most closely related to the previously reported SGCA strain, SGCA5. and JavaScript. Here we compare sequence capture and amplicon-based methods for sequencing SARS-CoV-2 and describe a streamlined tailed amplicon method for cost-effective and highly scalable SARS-CoV-2 sequencing. Get what matters in translational research, free to your inbox weekly. Bioinformatics. Nat Biotechnol. Di Paola N, Sanchez-Lockhart M, Zeng X, Kuhn JH, Palacios G. Viral genomics in Ebola virus research. Despite observing negligible amounts of primer dimer products on the bioanalyzer trace, samples with N1 and N2 Ct values greater than 30 had as much as 50% primer dimer in the resulting sequencing reads. For samples with Ct vales of less than 30, average coverage was 98.81% (10x) and 94.72% (100x) at a subsampled read depth of 100,000 raw reads (Fig. Google Scholar. The following reaction was set up for non-fragmented priming of RNA: 5L template RNA and 1L NEBNext Random Primers were combined and incubated at 65C for 5min. Provided by the Springer Nature SharedIt content-sharing initiative. More posts you may like r/labrats Join 9 days ago Lab archetypes 512 131 r/labrats Join 28 days ago Li, W., Hartung, J. S. & Levy, L. Quantitative real-time PCR for detection and identification of Candidatus Liberibacter species associated with citrus huanglongbing. Part of Bioinformatics. A) Percentage of genome coverage at 10x at different subsampled read depths for the indicated sample when sequenced using the indicated workflow. We estimated phylogenies of all samples along with 11 available reference genomes, using both a SNP and pan-genome approach. Targeted DNA enrichment and whole genome sequencing of Neisseria meningitidis directly from clinical specimens. Target-enrichment strategies for next-generation sequencing. An amplicon-based sequencing framework for accurately measuring intrahost virus diversity using PrimalSeq and iVar. Reference prophage genome sequences were at the top. Next, we assessed how well enrichment captures the genome diversity of different strains. S8. Each LHCA sample contained prophages SC1 and SC2, while SGCA samples contained only SC1 (Fig. Importantly, the RNA probe design of this positive capture method ensures retention of strain diversity, which other positive selection methods using primers run a risk of losing. Genome Announc, https://doi.org/10.1128/genomeA.00999-14 (2014). More importantly, this method significantly pushes the sequencing limitation to much lower titer samples while preserving strain diversity. A new coronavirus associated with human respiratory disease in China. Manufacturer: Agilent - Keysight. The tailed amplicon approach we describe bypasses costly and labor-intensive library preparation steps and will allow for production of SARS-CoV-2 libraries at high scale (similar workflows are run on tens of thousands of samples per year in the University of Minnesota Genomics Center) at low cost (between $2040 per sample depending on scale, including labor costs). An alternative to the Agilent Bioanalyzer is Biorads Experion system. $12,500 USD. S7. Emerg Infect Dis. We have previously reported a substantial size bias on the MiSeq, which may help explain the preferential clustering and out-sized proportion of primer dimer reads present in the sequencing data for some samples [16]. Percentage of genome coverage at 10x at different subsampled read depths for WA1 and UMGC SARS-CoV-2 isolates sequenced with different methods. Mol Plant Microbe Interact. Cornell Visualization and Imaging Partnership, Ask Us Anything About Your Needs or Projects. Candidatus Liberibacter americanus, associated with citrus huanglongbing (greening disease) in So Paulo State, Brazil. The BEI WA1 isolate strain was amplified for both 25 or 35 PCR cycles, using the same enzymes and PCR conditions used for the ARTIC v3 data set. By submitting a comment you agree to abide by our Terms and Community Guidelines. The analysis method for amplicon libraries is as follows: Sample quality was assessed with FastQC [19]. Bioinformatics. 31(22), 36913693 (2015). Percentage of genome coverage at 100x at different subsampled read depths for WA1 and UMGC SARS-CoV-2 isolates sequenced with different methods. An estimated 10,000 viral genome copies were used as input for cDNA generation. Advanced Analytical is my personal favorite. M.S. Are there any alternatives to this that anyone can recommend that is more modern tech? How to Determine the DV200 of FFPE RNA Samples on the Agilent TapeStation This Information Applies To: 4200, 4150 and 2200 TapeStation, TapeStation analysis software A02.02 or higher. Mamanova, L. et al. All raw read files were deposited to the SRA public database under BioProject ID PRJNA540608. More importantly, this method also effectively captures regions of diversity in the CLas genome, which provides precise molecular characterization of different strains. Reads were discarded with a mean quality score of less than 10 or when shorter than 200 base pairs, to avoid potential probe contamination, using BBDuk v38.12 (http://bbtools.jgi.doe.gov). Successful grafted citrus trees were determined by HLBaspr real-time quantitative PCR from symptomatic leaves. Measuring sequencer size bias using REcount: a novel method for highly accurate Illumina sequencing-based quantification. If you have a disability and are having trouble accessing information on this website or need materials in an alternate format, contact [email protected] for assistance. S.N. The DV 200 score is a quality score for evaluating quality of RNA derived from formalin-fixed paraffin-embedded (FFPE) samples established by Illumina Inc. in 2016. . bioRxiv. b Percent of the BEI WA1 isolate genome coverage at 100x at different subsampled read depths when sequenced with the indicated approach. Percentage of genome coverage at 10x at different subsampled read depths for WA1 and UMGC SARS-CoV-2 isolates sequenced the tailed amplicon v1 method amplified for 25 PCR cycles in the first PCR reaction. It is difficult to draw a comparison between alternative methods since only a few multiplex amplicon-based papers to sequence the whole SARS-CoV-2 genome have . statement and 25(15), 19681969 (2009). Nat Methods. Click on the hotspots and explore videos, literature, and more! (b) SGCA samples at different Cq values: Cq 20 (blue), Cq 22 (red). 2017;12:12616. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The hybridized . Next, 1g of each library was hybridized with the SureSelect capture library. 2a-b, Supplemental Table1, Supplemental Table2). Therefore, it could be possible to obtain the whole genome with even lower titer if more reads are used for the sample. Number of total reads generated per sample using the Illumina Nextera DNA Flex Enrichment workflow relative to: A) Sample N1 Ct value; B) Sample N2 Ct value. For the ARTIC v3 protocol, the average coverage at a subsampled read depth of 100,000 raw reads was 98.97% (10x) and 95.14% (100x) for all five test samples. We thank Brandon Vanderbush for conducting QC on the SARS-CoV-2 samples and sequencing libraries. Hundreds of millions of sequencing reads are needed to get good coverage of CLas from an HLB positive citrus sample. The average coverage at a subsampled read depth of 100,000 raw reads was 99.89% (10x) and 75.90% (100x) for all six test samples (Supplemental Table1, Supplemental Table2). The following recipe was used to set up the PCR reactions: 2.5L template cDNA, 14.75L nuclease-free water, 5L 5x Q5 reaction buffer (New England Biolabs, Ipswich, MA), 0.5L 10mM dNTPs (Kapa Biosystems, Woburn, MA), 0.25L Q5 Polymerase (New England Biolabs, Ipswich, MA), 2L primer pool 1 or 2 (10M) for the tailed v1 protocol. Supplemental Fig. Samples are colored as in panels c-f. b Evenness of representation of amplicons for different workflows as a function of sample N1 Ct value. Supported on their Sequel II and IIe instruments, and now expanded to their latest Revio sequencer, HiFi sequencing is built, Long-read technologies have repeatedly demonstrated their value in genomics research. Methods for SARS-CoV-2 genome sequencing compared in this study. 3b, Supplemental Fig. Consistent with other recent analyses of SARS-CoV-2 amplicon sequencing approaches [17], we observed highly concordant results from samples with N1 and N2 Ct values of less than 30. Cookies policy. Proceedings of the 2nd International Citrus Canker and Huanglongbing Research Workshop 2005, Orlando Florida, USA, p50 (2005). Loop-mediated isothermal amplification (LAMP) assay for specific and rapid detection of Dickeya fangzhongdai targeting a unique genomic region, Nucleic acids enrichment of fungal pathogens to study host-pathogen interactions, Rapid Detection of Genetic Engineering, Structural Variation, and Antimicrobial Resistance Markers in Bacterial Biothreat Pathogens by Nanopore Sequencing, Multiple internal controls enhance reliability for PCR and real time PCR detection of Rathayibacter toxicus, Targeted enrichment outperforms other enrichment techniques and enables more multi-species RNA-Seq analyses, Metagenomic sequencing for detection and identification of the boxwood blight pathogen Calonectria pseudonaviculata, De novo assembly and annotation of three Leptosphaeria genomes using Oxford Nanopore MinION sequencing, Evaluation of Oxford Nanopores MinION Sequencing Device for Microbial Whole Genome Sequencing Applications, Critical steps in clinical shotgun metagenomics for the concomitant detection and typing of microbial pathogens, https://www.aphis.usda.gov/plant_health/plant_pest_info/citrus_greening/downloads/pdf_files/nationalquarantinemap.pdf, http://tree.bio.ed.ac.uk/software/figtree/, https://doi.org/10.1094/PHP-2007-0906-01-RV, https://doi.org/10.1371/journal.pone.0112968, https://doi.org/10.1094/PHYTO-08-17-0282-R, https://doi.org/10.1094/PHYTO-06-18-0185-R, http://creativecommons.org/licenses/by/4.0/, Sign up for Nature Briefing: Translational Research. Reads that did not align to the host genome were aligned to the reference Wuhan-Hu-1 [5] SARS-CoV-2 genome (MN908947.3) using BWA [21]. Sci Rep 9, 18962 (2019). Briefly, three separate 10L RT-qPCR reactions were set up in a 384-well Barcoded plate (Thermo Fisher Scientific, Waltham, MA) for either the N1, N2, or RP primers and probes. This allows functional sequencing libraries to be created through a second indexing PCR reaction that adds sample-specific barcodes and flow cell adapters. TapeStation Systems An Interactive Lab Experience, Chromatography & Spectroscopy Lab Supplies, GC Calculators & Method Translation Software, BioCalculators / Nucleic Acid Calculators.