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SRX20030753: GSM7196446: P20_9 [Z20_B2_1_S57]; Danio rerio; RNA-Seq
1 ILLUMINA (NextSeq 2000) run: 4.1M spots, 210.2M bases, 68.9Mb downloads

External Id: GSM7196446_r1
Submitted by: Department of Neuroscience, Carleton University
Study: Identifying suitable experimental designs to determining differentially expressed genes and benchmark doses from transcriptomic dose-response studies in zebrafish embryos
show Abstracthide Abstract
To identify suitable experimental designsfor toxicology experiments (i.e., sample size, pool size, number of biological replicates, and number of doses), we exposed zebrafish embryos to 0.1% dimethyl sulfoxide for 4-120 hours post fertilization. We then created 96 samples to examine the impact of pool size comprised of 1, 5, 10, and 20 zebrafish embryos each (n = 24). From these, total RNA extraction was extracted and transcriptomic analyses were done using the TempO-Seq S1500+ Surrogate Assay platform. Samples were sequencing on an Illumina NextSeq 2000. The data were simulated to conduct power analyses exploring suitable study designs for identification of differentially expressed genes and for benchmark dose analysis. The aim was to identify a feasible pool size and sample size that would yield an acceptable false discovery rate for these analyses. Overall design: Samples were comprised of different pool sizes of embryos (i.e., 1, 5, 10, and 20 embryos per sample, indicated by Z1, Z5, Z10, and Z20) to identify optimal experimental designs for the dose-response study. Each pool size included 24 samples and a total of 96 samples were sequenced on the TempO-Seq S1500+ Surrogate assay platform. This study consisted of 12 extraction batches with 8 samples per batch and 3 batches per pool size.
Sample: P20_9 [Z20_B2_1_S57]
SAMN34272252 • SRS17370326 • All experiments • All runs
Organism: Danio rerio
Library:
Name: GSM7196446
Instrument: NextSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNAs were extracted using a Qiagen Mini kit (Qiagen, Toronto, Canada), followed by the manufacter's protocol RNA libraries were prepared in a 96-sample TempO-Seq S1500+ Zebrafish Surrogate Assay panel (BioSpyder Technologies, California, USA). Each sample was diluted with an equal volume of 2X Enhanced Lysis Buffer. All samples were annealed with detector oligo probes and sequentially processed in digestion, ligation, and amplification steps according to the manufacturer's protocol. Equal amounts of each sample's libraries were pooled in a 1.5 ml microtube and purified using NucleoSpin® Gel and PCR Cleanup Kits (Macherey-Nagel, Düren, Germany). Purified pooled libraries were sequenced on an Illumina NextSeq 2000 using a P2 Reagent kit and a 100 cycle flow cell (Illumina, USA). TempO-Seq S1500+ Zebrafish Surrogate Assay - Index E set was used
Runs: 1 run, 4.1M spots, 210.2M bases, 68.9Mb
Run# of Spots# of BasesSizePublished
SRR242345614,122,515210.2M68.9Mb2024-04-17

ID:
27429489

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