
==== Front
Wellcome Open Res
Wellcome Open Res
Wellcome Open Research
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10.12688/wellcomeopenres.22272.1
Data Note
Articles
The genome sequence of a cased caddisfly, Mystacides longicornis (Linnaeus, 1758)
[version 1; peer review: 2 approved]

Coleman Derek Investigation Resources Writing – Original Draft Preparation Writing – Review & Editing 1
Natural History Museum Genome Acquisition Lab
Darwin Tree of Life Barcoding collective
Wellcome Sanger Institute Tree of Life Management, Samples and Laboratory team
Wellcome Sanger Institute Scientific Operations: Sequencing Operations
Wellcome Sanger Institute Tree of Life Core Informatics team
Tree of Life Core Informatics collective
Darwin Tree of Life Consortiuma
1 Independent researcher, Carshalton, England, UK
a mark.blaxter@sanger.ac.uk
No competing interests were disclosed.

13 6 2024
2024
9 31015 5 2024
Copyright: © 2024 Coleman D et al.
2024
https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

We present a genome assembly from an individual male Mystacides longicornis (cased caddisfly; Arthropoda; Insecta; Trichoptera; Leptoceridae). The genome sequence is 665.1 megabases in span. Most of the assembly is scaffolded into 20 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 15.75 kilobases in length.

Mystacides longicornis
cased caddisfly
genome sequence
chromosomal
Trichoptera
Wellcome Trust218328 206194 This work was supported by Wellcome through core funding to the Wellcome Sanger Institute [206194, <a href=https://doi.org/10.35802/206194>https://doi.org/10.35802/206194</a>] and the Darwin Tree of Life Discretionary Award [218328, <a href=https://doi.org/10.35802/218328>https://doi.org/10.35802/218328 </a>]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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pmcSpecies taxonomy

Eukaryota; Opisthokonta; Metazoa; Eumetazoa; Bilateria; Protostomia; Ecdysozoa; Panarthropoda; Arthropoda; Mandibulata; Pancrustacea; Hexapoda; Insecta; Dicondylia; Pterygota; Neoptera; Endopterygota; Amphiesmenoptera; Trichoptera; Integripalpia; Brevitentoria; Leptoceroidea; Leptoceridae; Leptocerinae; Mystacidini; Mystacides; Mystacides longicornis (Linnaeus, 1758) (NCBI:txid699848).

Background

Mystacides longicornis is a cased caddis in the family Leptoceridae, which are easily recognised by their very long antennae. The commonest form of the adult is also easily recognised, having wings with dark bands on a fawn background; the other form has plain brown wings. Other features making identification easy are the red eyes and black hairy maxillary palps that protrude outwards ( Barnard & Ross, 2012). The larval case is straight or slightly curved, composed of sand grains and plant fragments. Often fragments of the case project beyond the case, presumably to make them less easy for predators to swallow. The larva is omnivorous and can be found in both still and flowing water with the adults flying from May to September ( Wallace et al., 2003). It is widespread and very common In England and Wales but less so in Scotland ( NBN Atlas Partnership, 2024). It is widespread in Europe ( O’Connor, 2015).

The genome of Mystacides longicornis was sequenced as part of the Darwin Tree of Life Project, a collaborative effort to sequence all named eukaryotic species in the Atlantic Archipelago of Britain and Ireland. Here we present a chromosomal-level whole genome sequence for Mystacides longicornis, based on one male specimen from Lea Broad, England, UK.

Genome sequence report

The genome was sequenced from a male Mystacides longicornis ( Figure 1) collected from Lea Broad, England, UK (52.62, 1.23). A total of 43-fold coverage in Pacific Biosciences single-molecule HiFi long reads was generated. Primary assembly contigs were scaffolded with chromosome conformation Hi-C data. Manual assembly curation corrected 13 missing joins or mis-joins and removed 2 haplotypic duplications, reducing the scaffold number by 13.64%, and increasing the scaffold N50 by 0.34%.

Figure 1. Photograph of the Mystacides longicornis (iiMysLong1) specimen used for genome sequencing.

The final assembly has a total length of 665.1 Mb in 37 sequence scaffolds with a scaffold N50 of 33.6 Mb ( Table 1). The snail plot in Figure 2 provides a summary of the assembly statistics, while the distribution of assembly scaffolds on GC proportion and coverage is shown in Figure 3. The cumulative assembly plot in Figure 4 shows curves for subsets of scaffolds assigned to different phyla. Most (99.88%) of the assembly sequence was assigned to 20 chromosomal-level scaffolds, representing 19 autosomes and the Z sex chromosome. Chromosome-scale scaffolds confirmed by the Hi-C data are named in order of size ( Figure 5; Table 2). Chromosome Z was assigned based on synteny to Athripsodes cinereus (GCA_947579605.1) ( Wallace et al., 2023). While not fully phased, the assembly deposited is of one haplotype. Contigs corresponding to the second haplotype have also been deposited. The mitochondrial genome was also assembled and can be found as a contig within the multifasta file of the genome submission.

Table 1. Genome data for Mystacides longicornis, iiMysLong1.1.

Project accession data	
Assembly identifier	iiMysLong1.1	
Species	Mystacides longicornis	
Specimen	iiMysLong1	
NCBI taxonomy ID	699848	
BioProject	PRJEB66746	
BioSample ID	SAMEA112964140	
Isolate information	iiMysLong1, whole organism (PacBio DNA and
Illumina Hi-C sequencing)	
Assembly metrics *	Benchmark	
Consensus quality (QV)	67.1	≥ 50	
k-mer completeness	100.0%	≥ 95%	
BUSCO **	C:96.0%[S:95.1%,D:0.9%],
F:2.4%,M:1.6%,n:2,124	C ≥ 95%	
Percentage of assembly
mapped to chromosomes	99.88%	≥ 95%	
Sex chromosomes	Z	localised homologous pairs	
Organelles	Mitochondrial genome:
15.75 kb	complete single alleles	
Raw data accessions	
PacificBiosciences Sequel IIe	ERR12102452	
Hi-C Illumina	ERR12102418	
Genome assembly	
Assembly accession	GCA_963576905.1	
Accession of alternate haplotype	GCA_963576805.1	
Span (Mb)	665.1	
Number of contigs	246	
Contig N50 length (Mb)	5.1	
Number of scaffolds	37	
Scaffold N50 length (Mb)	33.6	
Longest scaffold (Mb)	41.43	
* Assembly metric benchmarks are adapted from column VGP-2020 of “Table 1: Proposed standards and metrics for defining genome assembly quality” from Rhie et al. (2021).

** BUSCO scores based on the endopterygota_odb10 BUSCO set using version v5.4.3. C = complete [S = single copy, D = duplicated], F = fragmented, M = missing, n = number of orthologues in comparison. A full set of BUSCO scores is available at https://blobtoolkit.genomehubs.org/view/Mystacides_longicornis/dataset/GCA_963576905.1/busco.

Figure 2. Genome assembly of Mystacides longicornis, iiMysLong1.1: metrics.

The BlobToolKit snail plot shows N50 metrics and BUSCO gene completeness. The main plot is divided into 1,000 size-ordered bins around the circumference with each bin representing 0.1% of the 665,109,236 bp assembly. The distribution of sequence lengths is shown in dark grey with the plot radius scaled to the longest sequence present in the assembly (41,430,793 bp, shown in red). . Orange and pale-orange arcs show the N50 and N90 sequence lengths (33,624,892 and 27,270,352 bp), respectively. The pale grey spiral shows the cumulative sequence count on a log scale with white scale lines showing successive orders of magnitude. The blue and pale-blue area around the outside of the plot shows the distribution of GC, AT and N percentages in the same bins as the inner plot. A summary of complete, fragmented, duplicated and missing BUSCO genes in the endopterygota_odb10 set is shown in the top right. An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/Mystacides_longicornis/dataset/GCA_963576905.1/snail.

Figure 3. Genome assembly of Mystacides longicornis, iiMysLong1.1: BlobToolKit GC-coverage plot.

Sequences are coloured by phylum. Circles are sized in proportion to sequence length. Histograms show the distribution of sequence length sum along each axis. An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/Mystacides_longicornis/dataset/GCA_963576905.1/blob.

Figure 4. Genome assembly of Mystacides longicornis, iiMysLong1.1: BlobToolKit cumulative sequence plot.

The grey line shows cumulative length for all sequences. Coloured lines show cumulative lengths of sequences assigned to each phylum using the buscogenes taxrule. An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/Mystacides_longicornis/dataset/GCA_963576905.1/cumulative.

Figure 5. Genome assembly of Mystacides longicornis, iiMysLong1.1: Hi-C contact map of the iiMysLong1.1 assembly, visualised using HiGlass.

Chromosomes are shown in order of size from left to right and top to bottom. An interactive version of this figure may be viewed at https://genome-note-higlass.tol.sanger.ac.uk/l/?d=Ndau4nLsQF2mSl7UaJ5hCA.

Table 2. Chromosomal pseudomolecules in the genome assembly of Mystacides longicornis, iiMysLong1.

INSDC accession	Chromosome	Length (Mb)	GC%	
OY756420.1	1	41.43	34.5	
OY756421.1	2	41.25	34.5	
OY756422.1	3	40.82	34.5	
OY756423.1	4	38.54	34.5	
OY756424.1	5	38.16	34.5	
OY756425.1	6	35.65	34.5	
OY756426.1	7	35.03	34.5	
OY756427.1	8	35.01	34.5	
OY756428.1	9	33.62	34.5	
OY756430.1	10	31.19	35.0	
OY756431.1	11	31.14	34.5	
OY756432.1	12	31.01	34.5	
OY756433.1	13	30.46	34.5	
OY756434.1	14	29.69	34.5	
OY756435.1	15	29.09	34.5	
OY756436.1	16	28.42	34.5	
OY756437.1	17	27.27	35.0	
OY756438.1	18	27.17	34.5	
OY756439.1	19	26.59	35.0	
OY756429.1	Z	32.82	34.5	
OY756440.1	MT	0.02	16.0	

The estimated Quality Value (QV) of the final assembly is 67.1 with k-mer completeness of 100.0%, and the assembly has a BUSCO v5.4.3 completeness of 96.0% (single = 95.1%, duplicated = 0.9%), using the endopterygota_odb10 reference set ( n = 2,124).

Metadata for specimens, BOLD barcode results, spectra estimates, sequencing runs, contaminants and pre-curation assembly statistics are given at https://links.tol.sanger.ac.uk/species/699848.

Methods

Sample acquisition and nucleic acid extraction

A male Mystacides longicornis (specimen ID NHMUK014438422, ToLID iiMysLong1) was hand-picked from Lea Broad, England, UK (latitude 52.62, longitude 1.23) on 2022-04-07. The specimen was hand-picked by Derek Coleman (Dipterists Forum), who also formally identified the species. It was preserved by dry-freezing at –80°C.

The workflow for high molecular weight (HMW) DNA extraction at the Wellcome Sanger Institute (WSI) Tree of Life Core Laboratory includes a sequence of core procedures: sample preparation; sample homogenisation, DNA extraction, fragmentation, and clean-up. The sample was prepared for DNA extraction at the WSI Tree of Life Core Laboratory: the iiMysLong1 sample was weighed and dissected on dry ice ( Jay et al., 2023). Tissue of the whole organism was homogenised using a PowerMasher II tissue disruptor ( Denton et al., 2023a), setting aside tissue for Hi-C sequencing.

HMW DNA was extracted in the WSI Scientific Operations core using the Automated MagAttract v2 protocol ( Oatley et al., 2023). The DNA was sheared into an average fragment size of 12–20 kb in a Megaruptor 3 system with speed setting 31 ( Bates et al., 2023). Sheared DNA was purified by solid-phase reversible immobilisation ( Strickland et al., 2023): in brief, the method employs a 1.8X ratio of AMPure PB beads to sample to eliminate shorter fragments and concentrate the DNA. The concentration of the sheared and purified DNA was assessed using a Nanodrop spectrophotometer and Qubit Fluorometer and Qubit dsDNA High Sensitivity Assay kit. Fragment size distribution was evaluated by running the sample on the FemtoPulse system.

Protocols developed by the WSI Tree of Life laboratory are publicly available on protocols.io ( Denton et al., 2023b).

Sequencing

Pacific Biosciences HiFi circular consensus DNA sequencing libraries were constructed according to the manufacturers’ instructions. DNA sequencing was performed by the Scientific Operations core at the WSI on a Pacific Biosciences Sequel IIe (HiFi) instrument. Hi-C data were also generated from remaining tissue of iiMysLong1 using the Arima2 kit and sequenced on the Illumina NovaSeq 6000 instrument.

Genome assembly and curation

Assembly was carried out with Hifiasm ( Cheng et al., 2021) and haplotypic duplication was identified and removed with purge_dups ( Guan et al., 2020). The assembly was then scaffolded with Hi-C data ( Rao et al., 2014) using YaHS ( Zhou et al., 2023). The assembly was checked for contamination and corrected using the TreeVal pipeline ( Pointon et al., 2023). Manual curation was performed using JBrowse2 ( Diesh et al., 2023), HiGlass ( Kerpedjiev et al., 2018) and PretextView ( Harry, 2022). The mitochondrial genome was assembled using MitoHiFi ( Uliano-Silva et al., 2023), which runs MitoFinder ( Allio et al., 2020) or MITOS ( Bernt et al., 2013) and uses these annotations to select the final mitochondrial contig and to ensure the general quality of the sequence.

Final assembly evaluation

The final assembly was post-processed and evaluated with the three Nextflow ( Di Tommaso et al., 2017) DSL2 pipelines “sanger-tol/readmapping” ( Surana et al., 2023a), “sanger-tol/genomenote” ( Surana et al., 2023b), and “sanger-tol/blobtoolkit” ( Muffato et al., 2024). The pipeline sanger-tol/readmapping aligns the Hi-C reads with bwa-mem2 ( Vasimuddin et al., 2019) and combines the alignment files with SAMtools ( Danecek et al., 2021). The sanger-tol/genomenote pipeline transforms the Hi-C alignments into a contact map with BEDTools ( Quinlan & Hall, 2010) and the Cooler tool suite ( Abdennur & Mirny, 2020), which is then visualised with HiGlass ( Kerpedjiev et al., 2018). It also provides statistics about the assembly with the NCBI datasets ( Sayers et al., 2024) report, computes k-mer completeness and QV consensus quality values with FastK and MerquryFK, and a completeness assessment with BUSCO ( Manni et al., 2021).

The sanger-tol/blobtoolkit pipeline is a Nextflow port of the previous Snakemake Blobtoolkit pipeline ( Challis et al., 2020). It aligns the PacBio reads with SAMtools and minimap2 ( Li, 2018) and generates coverage tracks for regions of fixed size. In parallel, it queries the GoaT database ( Challis et al., 2023) to identify all matching BUSCO lineages to run BUSCO ( Manni et al., 2021). For the three domain-level BUSCO lineage, the pipeline aligns the BUSCO genes to the Uniprot Reference Proteomes database ( Bateman et al., 2023) with DIAMOND ( Buchfink et al., 2021) blastp. The genome is also split into chunks according to the density of the BUSCO genes from the closest taxonomically lineage, and each chunk is aligned to the Uniprot Reference Proteomes database with DIAMOND blastx. Genome sequences that have no hit are then chunked with seqtk and aligned to the NT database with blastn ( Altschul et al., 1990). All those outputs are combined with the blobtools suite into a blobdir for visualisation.

All three pipelines were developed using the nf-core tooling ( Ewels et al., 2020), use MultiQC ( Ewels et al., 2016), and make extensive use of the Conda package manager, the Bioconda initiative ( Grüning et al., 2018), the Biocontainers infrastructure ( da Veiga Leprevost et al., 2017), and the Docker ( Merkel, 2014) and Singularity ( Kurtzer et al., 2017) containerisation solutions.

Table 3 contains a list of relevant software tool versions and sources.

Table 3. Software tools: versions and sources.

Software tool	Version	Source	
BEDTools	2.30.0	https://github.com/arq5x/bedtools2	
Blast	2.14.0	ftp://ftp.ncbi.nlm.nih.gov/blast/executables/blast+/	
BlobToolKit	4.3.7	https://github.com/blobtoolkit/blobtoolkit	
BUSCO	5.4.3 and 5.5.0	https://gitlab.com/ezlab/busco	
bwa-mem2	2.2.1	https://github.com/bwa-mem2/bwa-mem2	
Cooler	0.8.11	https://github.com/open2c/cooler	
DIAMOND	2.1.8	https://github.com/bbuchfink/diamond	
fasta_windows	0.2.4	https://github.com/tolkit/fasta_windows	
FastK	427104ea91c78c3b8b8b49f1a7d6bbeaa869ba1c	https://github.com/thegenemyers/FASTK	
GoaT CLI	0.2.5	https://github.com/genomehubs/goat-cli	
Hifiasm	0.19.5-r587	https://github.com/chhylp123/hifiasm	
HiGlass	44086069ee7d4d3f6f3f0012569789ec138f42b84a
a44357826c0b6753eb28de	https://github.com/higlass/higlass	
MerquryFK	d00d98157618f4e8d1a9190026b19b471055b22e	https://github.com/thegenemyers/MERQURY.FK	
MitoHiFi	3	https://github.com/marcelauliano/MitoHiFi	
MultiQC	1.14, 1.17, and 1.18	https://github.com/MultiQC/MultiQC	
NCBI Datasets	15.12.0	https://github.com/ncbi/datasets	
Nextflow	23.04.0-5857	https://github.com/nextflow-io/nextflow	
PretextView	0.2	https://github.com/wtsi-hpag/PretextView	
purge_dups	1.2.5	https://github.com/dfguan/purge_dups	
samtools	1.16.1, 1.17, and 1.18	https://github.com/samtools/samtools	
sanger-tol/genomenote	1.1.1	https://github.com/sanger-tol/genomenote	
sanger-tol/readmapping	1.2.1	https://github.com/sanger-tol/readmapping	
Seqtk	1.3	https://github.com/lh3/seqtk	
Singularity	3.9.0	https://github.com/sylabs/singularity	
TreeVal	1.0.0	https://github.com/sanger-tol/treeval	
YaHS	1.2a.2	https://github.com/c-zhou/yahs	

Wellcome Sanger Institute – Legal and Governance

The materials that have contributed to this genome note have been supplied by a Darwin Tree of Life Partner. The submission of materials by a Darwin Tree of Life Partner is subject to the ‘Darwin Tree of Life Project Sampling Code of Practice’, which can be found in full on the Darwin Tree of Life website here. By agreeing with and signing up to the Sampling Code of Practice, the Darwin Tree of Life Partner agrees they will meet the legal and ethical requirements and standards set out within this document in respect of all samples acquired for, and supplied to, the Darwin Tree of Life Project.

Further, the Wellcome Sanger Institute employs a process whereby due diligence is carried out proportionate to the nature of the materials themselves, and the circumstances under which they have been/are to be collected and provided for use. The purpose of this is to address and mitigate any potential legal and/or ethical implications of receipt and use of the materials as part of the research project, and to ensure that in doing so we align with best practice wherever possible. The overarching areas of consideration are:

•   Ethical review of provenance and sourcing of the material

•   Legality of collection, transfer and use (national and international)

Each transfer of samples is further undertaken according to a Research Collaboration Agreement or Material Transfer Agreement entered into by the Darwin Tree of Life Partner, Genome Research Limited (operating as the Wellcome Sanger Institute), and in some circumstances other Darwin Tree of Life collaborators.

Data availability

European Nucleotide Archive: Mystacides longicornis. Accession number PRJEB66746; https://identifiers.org/ena.embl/PRJEB66746 ( Wellcome Sanger Institute, 2023). The genome sequence is released openly for reuse. The Mystacides longicornis genome sequencing initiative is part of the Darwin Tree of Life (DToL) project. All raw sequence data and the assembly have been deposited in INSDC databases. The genome will be annotated using available RNA-Seq data and presented through the Ensembl pipeline at the European Bioinformatics Institute. Raw data and assembly accession identifiers are reported in Table 1.

Author information

Members of the Natural History Museum Genome Acquisition Lab are listed here: https://doi.org/10.5281/zenodo.7139035.

Members of the Darwin Tree of Life Barcoding collective are listed here: https://doi.org/10.5281/zenodo.4893703.

Members of the Wellcome Sanger Institute Tree of Life Management, Samples and Laboratory team are listed here: https://doi.org/10.5281/zenodo.10066175.

Members of Wellcome Sanger Institute Scientific Operations: Sequencing Operations are listed here: https://doi.org/10.5281/zenodo.10043364.

Members of the Wellcome Sanger Institute Tree of Life Core Informatics team are listed here: https://doi.org/10.5281/zenodo.10066637.

Members of the Tree of Life Core Informatics collective are listed here: https://doi.org/10.5281/zenodo.5013541.

Members of the Darwin Tree of Life Consortium are listed here: https://doi.org/10.5281/zenodo.4783558.

10.21956/wellcomeopenres.24549.r94734
Reviewer response for version 1
Cedden Doga 1Referee https://orcid.org/0000-0001-9031-3710

1 University of Göttingen, Göttingen, Germany
6 9 2024 Copyright: © 2024 Cedden D
2024
https://creativecommons.org/licenses/by/4.0/ This is an open access peer review report distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Version 1recommendationapprove
The data note by Derek Coleman presents a genome assembly from  Mystacides longicornis of sufficient quality.  The insect is adequately described, and the species taxonomy is accurate. The author presents a sufficient quality assessment of the assembly. The assembly was made publicly accessible. My only concern regarding the data note is that the antennae are not present in Figure 1 and should be one of the defining features of this species ( longicornis). Please consider replacing Figure 1. with a more appropriate photograph where the antennae are present and visible.

Are sufficient details of methods and materials provided to allow replication by others?

Yes

Is the rationale for creating the dataset(s) clearly described?

Yes

Are the datasets clearly presented in a useable and accessible format?

Yes

Are the protocols appropriate and is the work technically sound?

Yes

Reviewer Expertise:

I work on beetle pests using RNAi and transcriptomics.

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

10.21956/wellcomeopenres.24549.r94741
Reviewer response for version 1
Li Ruiqi 1Referee https://orcid.org/0000-0001-9428-6094

1 University of Colorado Boulder, Boulder, Colorado, USA
28 8 2024 Copyright: © 2024 Li R
2024
https://creativecommons.org/licenses/by/4.0/ This is an open access peer review report distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Version 1recommendationapprove
Coleman et al. have presented a high-quality genome of the cased caddisfly, Mystacides longicornis (Linnaeus, 1758). The study is well-conducted, and I have only a few minor suggestions for improvement.

1. Introduction: It would be beneficial to include a short paragraph on the potential applications of the genome. For example, are there any notable biological features within the clade whose genetic mechanisms are not yet understood? Additionally, how might this genome contribute to species delimitation and systematics within the group?

2. Figure 1: Adding a scale bar to Figure 1

3. Sample Acquisition: In the "Sample acquisition" section, I suggest combining the two sentences mentioning that the specimen “… was hand-picked from Lea Broad by Derek Coleman…” to avoid potential confusion.

3. Could you provide details on how the sample was prepared? Was the specimen snap-frozen, and if so, how was this process handled?

4. RNA Sequencing: If RNA was sequenced as part of this study, please include details on how it was extracted.

4. Did the authors verify the chromosome numbers? There are two other genomes available for Leptocerinae, both of which report 25 chromosomes (Wallace et al. 2023, Coleman et al. 2024). Typically, chromosome numbers are consistent among closely related species. For example, within the subfamily Tridacninae, most species have 18 chromosomes, with Tridacna gigas being a slight exception with 19 chromosomes, which is still very close to 18 (Li et al., 2024 3,4,5,6). It would be helpful to double check the numbers.

Are sufficient details of methods and materials provided to allow replication by others?

Yes

Is the rationale for creating the dataset(s) clearly described?

Yes

Are the datasets clearly presented in a useable and accessible format?

Yes

Are the protocols appropriate and is the work technically sound?

Yes

Reviewer Expertise:

Genomics

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

Competing interests: No competing interests were disclosed.

Competing interests: No competing interests were disclosed.
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