2025-10-30
ENA first public
ERC000011
ENA-CHECKLIST
UDE
INSDC center name
public
INSDC status
ERS25351942
SRA accession
9_G3
Submitter Id
Open Access. You are free to share and adapt under the following terms: You must give appropriate credit , provide a link to the license, and indicate if changes were made . You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.
accessRights
AATGATACGGCGACCACCGAGATCTACAC
adapter_forward
CAAGCAGAAGACGGCATACGAGAT
adapter_reverse
205
ampliconSize
10
amplificationReactionVolume
58
annealingTemp
fwh (megabarcoding)
assay_name
other: DNA megabarcoding - single specimen barcoding using high troughput sequencing
assay_type
Additional Image Data: DOI: 10.5281/zenodo.15966820
associated_resource
two-step PCR
barcoding_pcr_appr
Will be added upon release.
bibliographicCitation
1
checkls_ver
https://github.com/PhilippMRehsen/EPT-14_code
code_repo
2022
collection date
DreamTaq Green PCR Master Mix, Thermo Fisher Scientific
commercial_mm
0
demux_max_mismatch
Demultiplexer v1.2
demux_tool
Megabarcoding of single Insect (Ephemeroptera, Plecoptera, Trichoptera) specimen
description
Germany
geographic location (country and/or sea)
The extracts were tested using a serial dilusion approach (1:10, 1:50, 1:100, 1:200), followed by 40 cycles of PCR and visual evaluation via gel electrophoresis. In case undiluted samples didn't show visual bands after gel electrophoresis, samples were repeated using a 1:200 dilution.
inhibition_check
1
inhibition_check_0_1
University of Duisburg-Essen
institution
https://ror.org/04mz5ra38
institutionID
NovaSeq6000
instrument
paired end
lib_layout
https://creativecommons.org/licenses/by/4.0/
license
5
merge_min_overlap
APSCALE v1.7.1
merge_tool
195
min_len_cutoff
APSCALE v1.7.1
min_len_tool
2025-07-31 00:00:00
mod_date
1
neg_cont_0_1
insect metagenome
organism
98
otu_clust_cutoff
v2.22.1 (using APSCALE v1.7.1)
otu_clust_tool
BOLD
otu_db
BOLDigger v2.1.3
otu_seq_comp_appr
10
pcr2_amplificationReactionVolume
61
pcr2_annealingTemp
DreamTaq Green PCR Master Mix, Thermo Fisher Scientific
pcr2_commercial_mm
initial denaturation:95_3;denaturation:95_0.5;annealing:61_0.5;elongation:72_1;final elongation:72_10;10
pcr2_cond
10
pcr2_cycles
2
pcr2_dna_vol
1
pcr_0_1
initial denaturation:95_3;denaturation:95_0.5;annealing:58_1.5;elongation:72_1;final elongation:72_10;40
pcr_cond
40
pcr_cycles
1
pcr_dna_vol
10
pcr_primer_conc_forward
10
pcr_primer_conc_reverse
GGDACWGGWTGAACWGTWTAYCCHCC
pcr_primer_forward
fwhF2
pcr_primer_name_forward
fwhR2n
pcr_primer_name_reverse
https://doi.org/10.3897/mbmg.1.14625
pcr_primer_reference_forward
https://doi.org/10.3897/mbmg.1.14625
pcr_primer_reference_reverse
GTRATWGCHCCDGCTARWACWGG
pcr_primer_reverse
0.2
pcr_primer_vol_forward
0.2
pcr_primer_vol_reverse
0
pcr_rep
0
pos_cont_0_1
philipp.rehsen@uni-due.de
project_contact
exstream_germany_boye_2022_EPT14
project_id
Philipp M. Rehsen | Mia S. Honka | Mikko Impiö | Iris Madge Pimentel | Florian Leese | Arne J. Beermann
recordedBy
https://orcid.org/0009-0003-0156-457X | | https://orcid.org/0000-0001-7672-3000 | https://orcid.org/0000-0002-8252-7660 | https://orcid.org/0000-0002-5465-913X | https://orcid.org/0000-0003-0403-0322
recordedByID
Aquatic Ecosystem Research - University of Duisburg Essen, Germany
rightsHolder
insect metagenome
scientific_name
Taxa detected in negative controls were checked and labwork of samples on the same plate as negatice controls with high read numbers were re-done (starting with the extracts) and sequenced an additional time.
screen_contam_method
Samples returning non-target species (i.e., species not belonging to Ephemeroptera, Plecoptera or Trichoptera species) were removed in the creation of the final dataset
screen_nontarget_method
see corresponging paper
screen_other
NovaSeq6000 2 x 150 bp
seq_kit
Azenta Life Sciences (Leipzig, Germany)
sequencing_location
https://github.com/DominikBuchner/demultiplexer | https://github.com/DominikBuchner/apscale/ | https://github.com/DominikBuchner/BOLDigger3
sop_bioinformatics
Sterile labware kits used for each individual sample, all lab surfaces cleaned with 80% ethanol prior to extraction and PCR set up, single-use gloves were used during all steps in the lab.
sterilise_method
effect of multiple stressors (added salinity and increased temperature) on benthic macroinvetebrates (Ephemeroptera, Plecoptera, Trichoptera) in an outdoor field mesocosm experiment.
study_factor
invertebrate
targetTaxonomicAssay
invertebrate
targetTaxonomicScope
COI
target_gene
see fwhF2, fwhR2n primer
target_subfragment
sequence similarity
tax_assign_cat
98%: species level, 95%: genus level, 90%: family level, 85%: order level, <85%: class level
tax_class_collapse
75
tax_class_id_cutoff
75
tax_class_query_cutoff
Blue-Ray Biotech, TurboCycler
thermocycler
EPT-14_9_G3
title
cutadapt v4.4 (using APSCALE v1.7.1)
trim_method