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opencc-zeroNov 2022View details →

The topological nature of tag jumping in environmental DNA metabarcoding studies (sequencing raw data)

Metabarcoding of environmental DNA constitutes a state-of-the-art tool for environmental studies. One fundamental principle implicit in most metabarcoding studies is that individual sample amplicons can still be identified after being pooled with others – based on their unique combinations of tags – during the so-called demultiplexing step that follows sequencing. Nevertheless, it has been recognized that tags can sometimes be changed (i.e. tag jumping), which ultimately leads to sample crosstalk. Here, using four DNA metabarcoding datasets derived from the analysis of soils and sediments, we show that tag jumping follows very specific and systematic patterns. Specifically, we find a strong correlation between the number of reads in blank samples and their topological position in the tag matrix (described by vertical and horizontal vectors). This observed spatial pattern of artefactual sequences could be explained by polymerase activity, which leads to the exchange of the 3' tag of single stranded tagged sequences through the formation of heteroduplexes with mixed barcodes. Importantly, tag jumping substantially distorted our datasets – despite our use of methods suggested to minimize this error. We developed a topologic model to estimate the noise based on the counts in our blanks, which suggested that 40-80% of the taxa in our soil and sedimentary samples were likely false positives introduced through tag jumping. We highlight that the amount of false positive detections caused by tag jumping strongly biased our community analyses.

opencc-by-4.0Nov 2022View details →

Saved model and preprocessed data for "CRMnet:a deep learning model for predicting gene expression from large regulatory sequence datasets"

Saved TUNet model and preprocessed training data for "CRMnet: a deep learning model for predicting gene expression from large regulatory sequence datasets" To load the trained model: <pre><code class="language-python">import tensorflow as tf tf.keras.models.load_model("path to the model folder")</code></pre> for more information please find our repository: https://github.com/jiayuwen/CRMnet

opencc-by-4.0Dec 2022View details →

Appendix of the thesis "Acquisition of new genetic knowledge on strains of Mycobacterium bovis, circulating in France, by the whole genome sequencing approach." Ciriac CHARLES

Appendix 1 Table showing the Mycobacterium bovis genome information used in this study in panel 1 The 87 genomes are from previous work (Hauer et al., 2019). The names of the genomes are highlighted with color to distinguish them within a previously defined M. bovis cluster. In Yellow Cluster I/Clonal Complex Eu3. In orange Cluster G/Family F9. In brown Cluster C/Family SB0134. In purple Cluster D/Clonal Complex Eu1. In pink the Cluster A/Family F4. In blue the Cluster F/Clonal Complex Eu2. In grey, the others. Appendix 2 Table showing the Mycobacterium bovis genome information used in this study in panel 3. The 187 genomes are of genotypes F7 and F15 (SB0821 and SB0832 respectively) which are two closely related genotypes. These genomes had already been sequenced before the start of the thesis work. Some were published with the work of Duault and collaborators and others have not yet been published (noted NA) (Duault et al., 2022). Appendix 3 Table showing the Mycobacterium bovis genome information used in this study in panel 4. The 227 genomes are of genotype SB0120-DHV. These genomes had already been sequenced before the start of the thesis work but are currently unpublished (noted NA in the columns "Date of deposit" and "Genbank biosample"). Appendix 4 Position of IS6110 insertion sites in the panel of Mycobacterium bovis strains from Pyrénées-Atlantiques (SB0821 and SB0832). The position of the start and end of IS6110 are determined with the ISMapper tool and the reference genome AF2122/97. Appendix 5 Position of IS6110 insertion sites in the Mycobacterium bovis strain panel of SB0120-DHV. Position of the start and end of IS6110 are determined with the ISMapper tool and the Mb3601 reference genome. Appendix 6 This figure from the Modenesi internship report represents a circular consensus phylogenetic tree of 300 strains of SB0120-DHV (Modenesi 2019). The strains in panel 3 are part of these 300 strains and were selected according to their sequencing quality, which explains the smaller number of strains retained (227). This tree is divided into 11 different colored clades. The posterior probability values (which correspond to the probability that these clades are true) determined with Beast (Drummond and Rambaut, 2007) are represented inside the tree.

opencc-by-4.0Dec 2022View details →

Appendix of the thesis "Acquisition of new genetic knowledge on Mycobacterium bovis strains, circulating in France, by the whole genome sequencing approach", part "4. Sequencing of new complete genomes", Ciriac CHARLES

Annex S1: Sequencing metric of the 10 new genomes and obtained with fastqc. A: Metric provide to Illumina metric. B: metric provide to MinION metric. Annex S2: Pan-genomic study performed on 12 M. bovis complete genomes. The table indicates the genes accessory. “1” shows the presence of CDS and “0” his absence. Annex S3: Indels between the ten new complete genomes and Mb3601 using progressiveMauve. Annotation of these indels was performed with reference genome comparison for gap or with the annotation (with Prokka) of the new complete genome studied for insertion. IS6110 is marked in green. "Indel distribution" sheet shows the indel distribution on the M. bovis genome for the ten new complete genomes. Black arrows show the genomic region with the most of indel found. Annex S4: WgSNP analysis performed on 98 M. bovis. SNPs were annotated and selected according to their specificity to an M. bovis group described in Fig 5. The last common give information on the genetic impact of the SNP. The last sheet presents a graph of SNP number in genomic position. Annex S5: Alignments of the 12 M. bovis complete genomes.

Genome-wide RAD sequencing data suggest predominant role of vicariance in Sino-Japanese disjunction of the monotypic genus Conandron (Gesneriaceae)

Disjunct distribution is a key issue in biogeography and ecology, but it is often difficult to determine relative roles of dispersal vs. vicariance in disjunctions. We studied phylogeographic pattern of the monotypic Conandron ramondioides (Gesneriaceae), which shows Sino-Japanese disjunctions, with ddRAD sequencing based on a comprehensive sampling of 11 populations from mainland China, Taiwan Island, and Japan. We found a very high degree of genetic differentiation among these three regions, with very limited gene flow and a clear Isolation by Distance pattern. Mainland China and Japan clades diverged first from a widespread ancestral population in the middle Miocene, followed by a later divergence between mainland China and Taiwan Island clades in the early Pliocene. Three current groups have survived in various glacial refugia during the Last Glacial Maximum (LGM), and experienced contraction and/or bottlenecks since their divergence during Quaternary glacial cycles, with strong niche divergence between mainland China + Japan and Taiwan Island ranges. Thus, we verified a predominant role of vicariance in the current disjunction of the monotypic genus Conandron. The sharp phylogenetic separation, ecological niche divergences among these three groups and the great number of private alleles in all populations sampled indicate a considerable time of independent evolution and suggest the need for a taxonomic survey to detect potentially overlooked taxa.

CDR3 sequences of germ-free and conventional chickens

Microbial colonisation is paramount to the normal development of the immune system, particularly at mucosal sites. However, the relationships between the microbiome and the adaptive immune repertoire have mostly been explored in rodents and humans. Here, we report a high-throughput sequencing analysis of the chicken TCRβ repertoire and the influences of microbial colonisation on tissue-resident TCRβ+ cells. The results reveal that the microbiome is an important driver of TCRβ diversity in both intestinal tissues and the bursa of Fabricius, but not in the spleen. Of note, public TCRβ sequences (shared across individuals) make a substantial contribution to the repertoire. Additionally, different tissues exhibit biases in terms of their V family and J gene usage, and these effects were influenced by the gut-associated microbiome. TCRβ clonal expansions were identified in both colonised and germ-free birds, but differences between the groups were indicative of an influence of the microbiota. Together, these findings provide insight into the avian adaptive immune system and the influence of the microbiota on the TCRβ repertoire.

Flow cytometry YFP and CFP data and deep sequencing data of populations evolving in galactose

Copy-number and point mutations form the basis for most evolutionary novelty through the process of gene duplication and divergence. While a plethora of genomic sequence data reveals the long-term fate of diverging coding sequences and their cis-regulatory elements, little is known about the early dynamics around the duplication event itself. In microorganisms, selection for increased gene expression often drives the expansion of gene copy-number mutations, which serves as a crude adaptation, prior to divergence through refining point mutations. Using a simple synthetic genetic system that allows us to distinguish copy-number and point mutations, we study their early and transient adaptive dynamics in real-time in Escherichia coli. We find two qualitatively different routes of adaptation depending on the level of functional improvement selected for: In conditions of high gene expression demand, the two types of mutations occur as a combination. Under low gene expression demand, negative epistasis between the two types of mutations renders them mutually exclusive. Thus, owing to their higher frequency, adaptation is dominated by copy-number mutations. Ultimately, due to high rates of reversal and pleiotropic cost, copy-number mutations may not only serve as a crude and transient adaptation but also <a>constrain</a> sequence divergence over evolutionary time scales.

Data from: Phylogenomics of superrosids and core rosids based on nuclear sequences and synteny

Superrosids form one of the largest clades of angiosperms, including 18 orders (Vitales, Saxifragales and core rosids) which exhibits remarkable morphological and ecological diversity. However, phylogenetic relationships within superrosids remain unclear. To resolve the phylogeny of superrosids, we screened 122 single copy nuclear genes from 37 species, representing all 18 orders. Vitales was revealed as sister to all other superrosids. Within core rosids, the fabids should be restricted only to the nitrogen-fixing clade, while Picramniales, the CM clade, Huerteales, Oxalidales, Sapindales, Malvales and Brassicales composed an "expanded" malvids. The COM clade (sensu APG IV) did not form a monophyletic group. Crossosomatales, Geraniales, Myrtales and Zygophyllales did not belong to either malvids or fabids. The difficult phylogeny of superrosids is likely due to the combined effects of ancient reticulation and incomplete lineage sorting. To provide broader genomic representation of Saxifragales, we constructed a high-quality chromosome-level genome assembly for Tiarella polyphylla (Saxifragaceae). Whole genome microsynteny analysis of superrosids showed that Saxifragales shared more synteny clusters with core rosids than Vitales, which also indicated that Saxifragales has a closer relationship with core rosids. Our findings contribute to a better understanding of the phylogeny and evolution of angiosperms.

Solidago hybrid-sequence capture probe set

Premise of the study: The phylogenetic relationships among the ca. 138 species of goldenrods (Solidago; Asteraceae) have been difficult to infer due to species richness, and shallow interspecific genetic divergences. This study aims to overcome these obstacles by combining extensive sampling of goldenrod herbarium specimens with the use of a custom Solidago hybrid-sequence capture probe set. Methods: A set of tissues from herbarium samples comprising ca. 90% of Solidago species was assembled, and DNA was extracted. A custom hybrid-sequence capture probe set was designed, and data from 854 nuclear regions were obtained and analyzed from 209 specimens. Maximum likelihood and coalescent approaches were used to estimate the genus phylogeny for 157 diploid samples. Key results: Although DNAs from older specimens were both more fragmented and produced fewer sequencing reads, there was no relationship between specimen age and our ability to obtain sufficient data at the target loci. The Solidago phylogeny was generally well supported, with 88/155 (57%) nodes receiving ≥95% bootstrap support. Solidago was supported as monophyletic, with Chrysoma pauciflosculosa identified as sister. A clade comprising Solidago ericameriodes, Solidago odora, and Solidago chapmanii was identified as the earliest diverging Solidago lineage. The previously segregated genera Brintonia and Oligoneuron were identified as placed well within Solidago. These and other phylogenetic results were used to establish four subgenera and fifteen sections within the genus. Conclusions: The combination of expansive herbarium sampling and hybrid-sequence capture data allowed us to quickly and rigorously establish the evolutionary relationships within this difficult, species-rich group.

Population admixtures in medaka inferred by multiple arbitrary amplicon sequencing

Cost-effective genotyping can be achieved by sequencing PCR amplicons. Short 3–10 base primers can arbitrarily amplify thousands of loci using only a few primers. To improve the sequencing efficiency of the multiple arbitrary amplicon sequencing (MAAS) approach, we designed new primers and examined their efficiency in sequencing and genotyping. To demonstrate the effectiveness of our method, we applied it to examine the population structure of the small freshwater fish, medaka (Oryzias latipes). We obtained 2,987 informative SNVs with no missing genotype calls for 67 individuals from 15 wild populations and three artificial strains. The estimated phylogenic and population genetic structures of the wild populations were consistent with previous studies, corroborating the accuracy of our genotyping method. We also attempted to reconstruct the genetic backgrounds of a commercial orange mutant strain, Himedaka, which has caused a genetic disturbance in wild populations. Our admixture analysis focusing on Himedaka showed that at least two wild populations had genetically contributed to the nuclear genome of this mutant strain. Our genotyping methods and results will be useful in quantitative assessments of genetic disturbance by this commercially available strain.

The phylogeny and global biogeography of Primulaceae based on high-throughput DNA sequence data

The angiosperm family Primulaceae is morphologically diverse and distributed nearly worldwide. However, phylogenetic uncertainty has limited the ability to identify major morphological and biogeographic transitions. We used target capture sequencing with the Angiosperms353 kit for over 300 species across Ericales, tree-based sequence curation, and multiple phylogenetic approaches to investigate the phylogenetics of the major clades of Primulaceae and their relationship to other Ericales. The study included 150 samples of Primulaceae comprising nearly all recognized genera of the family, with a particular focus on the most diverse subfamily, Myrsinoideae, for which previous phylogenetic knowledge was poor. We used fossil and secondary calibrations to generate dated phylogenetic trees and conducted broad-scale biogeographic analyses as well as ancestral state reconstructions of plant habit.

Data from: Higher evolutionary dynamics of gene copy number for Drosophila glue genes located near short repeat sequences

Background During evolution, genes can experience duplications, losses, inversions and gene conversions. Why certain genes are more dynamic than others is poorly understood. Here we examine how several Sgs genes encoding glue proteins, which make up a bioadhesive that sticks the animal during metamorphosis, have evolved in Drosophila species. Results We examined high-quality genome assemblies of 24 Drosophila species to study the evolutionary dynamics of four glue genes that are present in D. melanogaster and are part of the same gene family – Sgs1, Sgs3, Sgs7 and Sgs8 – across approximately 30 millions of years. We annotated a total of 102 Sgs genes and grouped them into 4 subfamilies. We present here a new nomenclature for these Sgs genes based on protein sequence conservation, genomic location and presence/absence of internal repeats. Two types of glue genes were uncovered. The first category (Sgs1, Sgs3x, Sgs3e) showed a few gene losses but no duplication, no local inversion and no gene conversion. The second group (Sgs3b, Sgs7, Sgs8) exhibited multiple events of gene losses, gene duplications, local inversions and gene conversions. Our data suggest that the presence of short "new glue" genes near the genes of the latter group may have accelerated their dynamics. Conclusions Our comparative analysis suggests that the evolutionary dynamics of glue genes is influenced by genomic context. Our molecular, phylogenetic and comparative analysis of the four glue genes Sgs1, Sgs3, Sgs7 and Sgs8 provides the foundation for investigating the role of the various glue genes during Drosophila life.

opencc-by-4.0Oct 2022View details →

Datasets generated for KA-Search: rapid and exhaustive sequence identity search of known antibodies

Datasets generated for testing KA-Search in the paper "KA-Search: rapid and exhaustive sequence identity search of known antibodies". Includes OAS-test and the 100 randomly selected non-redundant heavy chains of therapeutics.

opencc-by-4.0Jan 2023View details →

B-other ALL classification by Targeted RNA-sequencing

We present a comprehensive genetic study of 144 pediatric B-other Acute Lymphoblastic Leukemia cases diagnosed and treated at Boldrini Children's Hospital (Brazil). We performed a targeted RNA-sequencing to evaluated the benefits of introducing genomic technologies into routine diagnostics. Targeted RNA-sequencing further classified 66.7% B-other cases. All 'classical' and novel ALL subgroups, except for iAMP21, hyper- and hypodiploid cases were identified. In addition, clinically important genetic alterations as druggable lesions and prognostic factors were found. Here, we uploaded files that contain:  1. Gene expression data from the entire cohort studied, n=184 (Log2+1 TPM normalized gene expression data). Tab-delimited file (.csv) that contains a row for gene, a column for each sample, and expression values for each gene in each sample.  Annotation labels are in the first three rows and in the first column. 2. A subset of 23 BAM (Binary Alignment/Map) files for representative ALL cases, including BCR-ABL1, ETV6-RUNX1, TCF3-PBX1, KMT2A-r, High-Hyperdiploidy; DUX4-r, iAMP21, PAX5-driven, ABL-class fusion, JAK2-fusion, EPOR-fusion, CRLF2-high, ZNF384-r, MEF2D-r, NUTM1-r, B-'rest', IKZF1del, and ERGdel. 3- Genetic information of the 23 BAM files provided (tab-delimited file, .txt).

opencc-by-4.0Jan 2023View details →

Single-cell RNA sequencing of mouse embryonic cells from the oocyte, 2-cell, 4-cell, 8-cell, blastocyst, and morula stages

STRT-N is a newly optimized single-cell RNA sequencing method for studies of early genome activation in mammalian preimplantation development. Single embryos from the oocyte, 2-cell, 4-cell, 8-cell, blastocyst, and morula stages were sampled for experiments and were sequenced using STRT-N method. Here is the raw data from STRTN-seq. FASTQ files are available in <a href="https://www.ebi.ac.uk/biostudies/studies/S-BSST976">BioStudies database</a>.

Chamaeleo africanus DNA sequencing purity results and statistics

Conservation of wildlife often depends on high quality molecular data to establish reliable species identification. Traditional approaches in extracting material (DNA) for phylogenetic studies on chameleons have relied on removed, euthanized or preserved/museum specimens, while field sampling usually takes the form of tail clippings from living individuals and their subsequent release. In this article, we propose an alternative to these approaches for field sampling, towards isolation of nuclear and mitochondrial DNA with oral (buccal) swabs, a methodology already been demonstrated as effective in other taxa. Options of sampling, storage, transport, extraction of DNA are presented and the quality and quantity of extracted material (using venipuncture as a positive control) was demonstrated as sufficient for downstream applications, including sequencing thereby presenting a practical field alternative. The advantages and limitations of this minimally invasive and non-destructive method applied to Chamaeleo africanus are further discussed.

16s rRNA sequencing of Wolbachia-infected and uninfected Drosophila male gut

Wolbachia are the most widely distributed intracellular bacteria, and their most common effect on host phenotype is cytoplasmic incompatibility (CI). A variety of models have been proposed to decipher the molecular mechanism of CI, in which the HM (host modification) model predicts that the effectors of Wolbachia play an important role in sperm modification. However, due to the complexity of spermatogenesis and cell-type heterogeneity in the testis, we still do not know whether Wolbachia have different effects on cells at different stages of spermatogenesis, nor whether these effects are linked with CI. Therefore, we used single-cell RNA sequencing to analyze gene expression profiles in the adult male Drosophila testes with or without Wolbachia infection. We found that Wolbachia significantly affected the proportion of different types of germ cells and affected multiple metabolic pathways in germ cells. Most importantly, Wolbachia had the greatest impact on germline stem cells (GSCs), resulting in the dysregulation expression of genes related to nucleosome assembly and CI, Wolbachia infection also influenced the histone-to-protamine transition in the late stage of sperm development. These results suggest that future studies of Wolbachia-mediated sperm modification should focus more on cells in the early stages of spermatogenesis.

Exome sequencing of a hybrid pine species complex on the Qinghai-Tibetan Plateau

This study investigates the evolutionary history of Pinus densata on the Qinghai-Tibetan Plateau (QTP) and genomic heterogeneity across a zone of species transition to understand contemporary dynamics of selection and evolution of species barriers. We analyzed the genetic diversity in a range-wide collection of P. densata and representative populations of its progenitors P. tabuliformis and P. yunnanensis using 40,000 exome probe capture sequencing.

opencc-by-4.0Dec 2022View details →

Genbank and BOLD accession numbers of sequences from "New insights into the diversity of cryptobenthic Cirripectes blennies in the Mascarene Archipelago sampled using Autonomous Reef Monitoring Structures (ARMS)"

Abstract: Autonomous Reef Monitoring Structures (ARMS) are artificial mini-reefs designed for standardised sampling of sessile and small motile cryptobenthic organisms. ARMS are also effective for collecting small cryptobenthic fishes, such as the combtooth blennies of the genus Cirripectes. Recent studies discovered several Cirripectes species endemic to islands or archipelagos, in spite of the generally broad distributions of tropical and subtropical blennies. Thus, in order to evaluate the diversity and distribution of Cirripectes species in the Mascarene Archipelago, a little studied region but an important biodiversity hotspot, complete mitochondrial genomes and nuclear rhodopsin genes were sequenced for 39 specimens collected with ARMS deployed on outer reef slopes at Reunion and Rodrigues islands. Mitochondrial COI sequences were analysed to integrate these specimens within the largest dataset of publicly available sequences. Three species were found in the Mascarene Archipelago, Cirripectes castaneus, C. randalli, and C. stigmaticus. C. castaneus and C. stigmaticus both have an Indo-Pacific distribution with several haplotypes shared among distant localities. In agreement with the literature, C. randalli shows a small-range endemism restricted to the Mascarenes. We confirmed the presence of C. castaneus, C. randalli and C. stigmaticus in Rodrigues, and the presence of C. stigmaticus in Reunion. This study contributes to fill the gaps in taxonomic and molecular knowledge of the reef cryptobiome in the South-West Indian Ocean, and provides the first complete mitogenomes for the genus, a crucial step for future molecular-based inventories (e.g. eDNA). Content of this repository: Genbank and BOLD accession numbers of sequences from "New insights into the diversity of cryptobenthic Cirripectes blennies in the Mascarene Archipelago sampled using Autonomous Reef Monitoring Structures (ARMS)" published in Ecology and Evolution (2023)