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Dataset results
18 results for “Direct PCR”
Data from: Drop it all: Extraction-free detection of non-indigenous marine species through optimized direct-droplet digital PCR
<p>Molecular biosecurity surveillance programs increasingly use environmental DNA (eDNA) for detecting marine non-indigenous species (NIS). However, the current molecular detection workflow is cumbersome, prone to errors and delays, and is limited in providing knowledge about eDNA beyond the spatial and temporal extent of the sampling. These limitations can hinder management efforts and restrict the "opportunity window" for a rapid response to new marine NIS incursions. Emerging innovative field-deployable digital droplet PCR (ddPCR) systems offer improved workflow efficiency by autonomously analyzing targeted free-floating extra-cellular eDNA (free-eDNA) signals. Despite their potential, these systems have not been tested in marine environments. Thus, an aquarium study was conducted with three distinct marine NIS: <span>the Mediterranean fanworm <em>Sabella spallanzanii</em>, the ascidian clubbed tunicate <em>Styela clava</em>, and the brown bryozoan <em>Bugula neritina</em></span> to evaluate the detectability of free-eDNA in seawater. The detectability of targeted free-eDNA was assessed by directly analyzing aquarium water samples using an optimized species-specific ddPCR assay, without filtration or DNA extraction, so-called, "direct-ddPCR". The results demonstrated the consistent detection of <em>Sabella spallanzanii</em> and <em>Bugula neritina</em> free-eDNA when these organisms were present in high abundance. Once organisms were removed, the free-eDNA signal exponentially declined, noting that free-eDNA persisted between 24-72 hours. Results indicate that organism biomass, specimen characteristics (e.g., stress and viability), and species-specific biological differences may influence free-eDNA detectability. These results are critical for implementing <em>in-situ</em> nucleic acid automated continuous sensing systems for marine biosurveillance, enabling point-of-need detection and <span>rapid management response to biosecurity threats. </span></p>
Drop it all: Extraction-free detection of non-indigenous marine species through optimized direct-droplet digital PCR
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Data from: Genotyping HapSTR loci: phase determination from direct sequencing of PCR products
HapSTRs combine information from a microsatellite (or simple tandem repeat, STR) with one or more single nucleotide polymorphisms (SNPs) in the DNA sequence immediately flanking the STR. These loci may offer increased power for the estimation of demographic parameters, but also present some challenges for data collection and analysis. We describe a process for inferring HapSTR alleles, including the flanking haplotypes, STR alleles, and their phase relative to each other, directly from DNA sequence electropherograms of PCR products from heterozygous individuals. Our approach eliminates the need for more costly and time-consuming processes such as cloning or acrylamide gel electrophoresis to separate alleles prior to sequencing.
Data from: “Direct PCR” optimization yields a rapid, cost-effective, non-destructive, and efficient method for obtaining DNA barcodes without DNA extraction
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Data from: Genotyping HapSTR loci: phase determination from direct sequencing of PCR products
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Data from: Trace DNA from insect skins: a comparison of five extraction protocols and direct PCR on chironomid pupal exuviae
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Improving results from touch DNA evidence with optimized direct PCR methods
<p>Improved methods to generate high-quality DNA profiles from touch DNA samples are of considerable interest to forensic DNA laboratories. Direct polymerase chain reaction (PCR) amplification, a sample processing method in which samples are added directly to amplification reactions without prior purification or quantification, has been identified as a method that may improve the generation of genotyping data from such samples; however, laboratories in the United States are required to use standard DNA processing methods to process low-level sample types. In part, this is due to FBI Quality Assurance Standard (QAS) 9.4, which requires all unknown forensic samples to undergo human-specific DNA quantification prior to amplification of short tandem repeat (STR) loci. The goal of this 3-year study was to generate data in support of a reevaluation of QAS 9.4.</p> <p>In two phases, this study examined the following: direct PCR-compatible collection methods in conjunction with mock touch DNA evidence samples on a variety of substrates (Phase I), direct PCR of touch DNA samples that were stored at room temperature for up to six months after collection with the optimum methods identified in Phase I, and direct PCR of touch DNA samples that were re-sampled after initial processing. Nine collection methods and various substrates were examined in Phase I, and three time points were examined in Phase II. In both phases, two processing methods were used: standard processing samples were extracted, quantified, and amplified in accordance with the QAS; and direct PCR samples were directly amplified. All STR profiles were assessed for overall profile quality.</p> <p>This project was supported by Award No. 2019-DU-BX-0009, awarded by the National Institute of Justice, Office of Justice Programs, U.S. Department of Justice. The opinions, findings, and conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect those of the Department of Justice.</p>
Supplementary material 1 from: Röder N, Schwenk K (2023) Direct PCR meets high-throughput sequencing – metabarcoding of chironomid communities without DNA extraction. Metabarcoding and Metagenomics 7: e102455. https://doi.org/10.3897/mbmg.7.102455
Overview of chironomid size classes
Supplementary material 2 from: Röder N, Schwenk K (2023) Direct PCR meets high-throughput sequencing – metabarcoding of chironomid communities without DNA extraction. Metabarcoding and Metagenomics 7: e102455. https://doi.org/10.3897/mbmg.7.102455
Composition of the two artificial chironomid communities
Routine Application of Point-of-care PCR Test to Identify and Direct Therapy for Acute Respiratory Infection in the Emergency Department Trial
ClinicalTrials.gov study NCT06780566. IPD Sharing: UNDECIDED. Countries: 1. Publications: 0.
Improving results from touch DNA evidence with optimized direct PCR methods
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Data from: Direct sequencing of haplotypes from diploid individuals through a modified emulsion PCR-based single-molecule sequencing approach
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Nanopore PCR-free direct cDNA sequencing of human kidney tissue
GEO Series GSE281264. Homo sapiens. 3 samples. Type: Expression profiling by high throughput sequencing.
Nanopore PCR-free direct cDNA sequencing on a murine model of endotoxin-indcued acute kidney injury
GEO Series GSE244942. Mus musculus. 17 samples. Type: Expression profiling by high throughput sequencing.
Data from: Direct PCR meets high-throughput sequencing - metabarcoding of chironomid communities without DNA extraction
<p>Abstract</p> <p>Metabarcoding is a valuable tool for investigating insect community compositions. However, high-throughput applications, such as for biomonitoring, require cost-effective and user-friendly procedures. To investigate if the time-consuming and labour-intensive DNA isolation step can be omitted in metabarcoding, we studied the difference in detection rates and individual read abundance using standard DNA isolation versus direct PCR protocols. Metabarcoding with and without DNA isolation was performed on artificially created communities with known composition as well as on natural communities both of the dipteran family Chironomidae to compare detection rates, individual read abundances and presence-absence community composition. The data sets include read abundances of all artificial and natural community samples. Compositions of the samples per data set are described in the respective README files. ASVs/OTUs and their respective DNA sequences are given. R Scripts for bioinformatic processing (dada2 for ASVs, JAMP for OTUs) are provided.</p> <p>Methods</p> <p>Chironomidae were retrieved from artificial ponds of the Eußerthal Ecosystem Research Station (EERES) near Landau, Germany, in 2019 and 2020. Adult specimens were collected from passive emergence traps. Chironomid samples were stored in 70% ethanol and later dried at 60°C. Samples were then finely ground using a bead mill. PCR-grade water was added to each tissue sample and thoroughly vortexed. The tissue-water mixes were frozen at -20°C until further analysis. Artificial communities were created by pipetting tissue-water mixes of individual chironomids. Natural communities from eight ponds and five consecutive sampling dates were selected to assess the applicability of the dPCR approach compared to standard metabarcoding protocols on natural chironomid communities. Four of the artificial ponds were treated with the mosquito control agent <em>Bacillus thuringiensis israelensis</em> (Bti). Tissue-water mixes of artificial and natural communities were both directly applied to PCR and used for DNA isolation. Illumina sequencing was performed and raw data were bioinformatically prepared. For more details see "Direct PCR meets high-throughput sequencing - metabarcoding of chironomid communities without DNA extraction" (Röder & Schwenk 2023). Raw sequences are available through GenBank SRA archive (BioProject accession number PRJNA989176). </p>
Detection of Azole Resistance Inducing Mutations on DNA Extracted Directly From Serum or Plasma of Immunocompromised Patients With an Invasive Aspergillus Infection Azole Resistance PCR Optimalization
ClinicalTrials.gov study NCT06069505. IPD Sharing: UNDECIDED. Countries: 1. Publications: 0.
Distinct 5′ and 3′ Coverage Biases Shape Transcriptome Interpretation in Nanopore Direct RNA versus PCR-cDNA Sequencing
GEO Series GSE306225. Homo sapiens. 12 samples. Type: Expression profiling by high throughput sequencing.
Real-time quantitative PCR analysis of human astrocytes exposed to pressure-driven flow and/or Direct Current Stimulation in vitro
GEO Series GSE207140. Homo sapiens. 140 samples. Type: Expression profiling by RT-PCR.
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These curated guides explain access requirements, typical timelines, costs, and reuse considerations for widely used research datasets.
Allen Brain Atlas
Allen Brain Atlas is an Allen Institute collection of brain map atlases, datasets, APIs, and analysis tools covering mouse, human, and non-human primate brain resources.
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DANDI Archive for NWB datasets
DANDI is a BRAIN Initiative archive for publishing and sharing neurophysiology data, including electrophysiology, optophysiology, and behavioral data packaged as NWB and related standards.
International Brain Laboratory public data
The International Brain Laboratory public data releases expose standardized mouse decision-making experiments, including Neuropixels recordings, widefield calcium imaging, behavior, and session metadata accessed through the ONE API.
OpenNeuro
OpenNeuro is a free, open platform for sharing neuroimaging datasets, with public search, dataset pages, and download paths for web, S3, DataLad, and the OpenNeuro CLI.