Principal investigators: Pr. Thomas Sauter (Luxembourg) / Pr. Dagmar Kulms (Dresden)
Experimenter: Sebastian Schindler (Dresden)
Sequencing on total RNA: Rashi Halder (Luxembourg)
24 FASTQ generated (deposited in ArrayExpress E-MTAB-10669)
10-1-Lasse_S10_R1_001.fastq.gz
10-3-Lasse_S16_R1_001.fastq.gz
11-1-Lasse_S11_R1_001.fastq.gz
11-3-Lasse_S17_R1_001.fastq.gz
1-1-Lasse_S1_R1_001.fastq.gz
12-1-Lasse_S12_R1_001.fastq.gz
12-3-Lasse_S18_R1_001.fastq.gz
1-3-Lasse_S7_R1_001.fastq.gz
2-1-Lasse_S2_R1_001.fastq.gz
2-3-Lasse_S8_R1_001.fastq.gz
3-1-Lasse_S3_R1_001.fastq.gz
3-3-Lasse_S9_R1_001.fastq.gz
4-1-Lasse_S4_R1_001.fastq.gz
4-3-Lasse_S10_R1_001.fastq.gz
5-1-Lasse_S5_R1_001.fastq.gz
5-3-Lasse_S11_R1_001.fastq.gz
6-1-Lasse_S6_R1_001.fastq.gz
6-3-Lasse_S12_R1_001.fastq.gz
7-1-Lasse_S7_R1_001.fastq.gz
7-3-Lasse_S13_R1_001.fastq.gz
8-1-Lasse_S8_R1_001.fastq.gz
8-3-Lasse_S14_R1_001.fastq.gz
9-1-Lasse_S9_R1_001.fastq.gz
9-3-Lasse_S15_R1_001.fastq.gz
The quality controls, read trimming, feature counting were performed using Snakemake and the tailor-made RNA-seq template.
The 3 config files are provided in the config
folder:
config.yaml
main parameterssamples.tsv
sample annotationsunits.tsv
link samples and FASTQ files
Main output is the DESeqDataSet
object from DESeq2
saved a binary file `deseq2/all.rds` after successful completion of the snakemake
pipeline.
See the file TRAIL_IZI.Rmd
, routines are loaded from R/utils.R
Performed on 1,248 genes using the BASH script run_genewalk.sh
on the https://hpc.uni.lu iris
machine.
The input file same_pdirection.txt
is also part of this repository.
The experiments presented in this paper were carried out using the HPC facilities of the University of Luxembourg (Varrette et al. 2014, https://hpc.uni.lu).
Varrette S., Bouvry P., Cartiaux H. and Georgatos F. Management of an Academic HPC Cluster: The UL Experience. Proc. of the 2014 Intl. Conf. on High Performance Computing & Simulation. HPCS 2014. 959-967.
software and algorithms | source | product number/identifier |
---|---|---|
AdapterRemoval | https://github.com/MikkelSchubert/adapterremoval | v2.3.1 |
apeglm | R bioconductor | v1.10.0 |
ComplexHeatmap | https://github.com/jokergoo/ComplexHeatmaps | v2.7.8.1000 |
DESeq2 | R bioconductor | v1.128.1 |
FALCON | De Landtsheer et al. 2017, https://github.com/sysbiolux/FALCON | |
GeneWalk | Ietswaart et al. 2021, https://churchman.med.harvard.edu/genewalk | v1.5.1 |
ggplot2 | R CRAN | v3.3.2 |
ggVennDiagram | R CRAN | v1.1 |
ggforce | R CRAN | v0.3.3 |
multiqc | https://multiqc.info/ | v1.9 |
python | https://python.org/ | v3.8.2 |
R Project for Statistical Computing | https://www.r-project.org | v4.0.0 |
RStudio | https://www.rstudio.com/ | v1.0.143 |
Rsubread | R bioconductor | v2.2.2 |
SAMtools | https://www.htslib.org | v1.10 |
Snakemake | https://snakemake.github.io | v5.20.1 |
STAR | https://github.com/alexdobin/STAR | v2.7.4a |
Tidyverse | R CRAN, http://tidyverse.org/ | v1.1.1 |
Tools citation
- Constantin Ahlmann-Eltze, Peter Hickey and Hervé Pagès (2021). MatrixGenerics: S4 Generic Smmary Statistic Functions that Operate on Matrix-Like Objects. R package version 1.4.0. htps://bioconductor.org/packages/MatrixGenerics - Gu, Z. (2016) Complex heatmaps reveal patterns and correlations in multidimensional genomic dta. Bioinformatics. - H. Pagès, M. Lawrence and P. Aboyoun (2021). S4Vectors: Foundation of vector-like and list-like cntainers in Bioconductor. R package version 0.30.0. https://bioconductor.org/packages/S4Vectors - H. Wickham. ggplot2: Elegant Graphics for Data Analysis. Springer-Verlag New York, 2016. - Hadley Wickham (2019). stringr: Simple, Consistent Wrappers for Common String Operations. R pckage version 1.4.0. https://CRAN.R-project.org/package=stringr - Hadley Wickham (2021). forcats: Tools for Working with Categorical Variables (Factors). R pckage version 0.5.1. https://CRAN.R-project.org/package=forcats - Hadley Wickham (2021). tidyr: Tidy Messy Data. R package version 1.1.3. htps://CRAN.R-project.org/package=tidyr - Hadley Wickham and Jim Hester (2020). readr: Read Rectangular Text Data. R package version 14.0. https://CRAN.R-project.org/package=readr - Hadley Wickham, Romain François, Lionel Henry and Kirill Müller (2021). dplyr: A Grammar of Data Mnipulation. R package version 1.0.7. https://CRAN.R-project.org/package=dplyr - Henrik Bengtsson (2021). matrixStats: Functions that Apply to Rows and Columns of Matrices (and t Vectors). R package version 0.59.0. https://CRAN.R-project.org/package=matrixStats - Jim Hester and Hadley Wickham (2021). vroom: Read and Write Rectangular Text Data Quickly. R pckage version 1.5.1. https://CRAN.R-project.org/package=vroom - Kamil Slowikowski (2021). ggrepel: Automatically Position Non-Overlapping Text Labels with 'gplot2'. R package version 0.9.1. https://CRAN.R-project.org/package=ggrepel - Kirill Müller and Hadley Wickham (2021). tibble: Simple Data Frames. R package version 3.1.2. htps://CRAN.R-project.org/package=tibble - Lawrence M, Huber W, Pag`es H, Aboyoun P, Carlson M, et al. (2013) Software for Computing and Anotating Genomic Ranges. PLoS Comput Biol 9(8): e1003118. doi:10.1371/journal.pcbi.1003118 - Lawrence M, Huber W, Pag`es H, Aboyoun P, Carlson M, et al. (2013) Software for Computing and Annotating Genomic Ranges. PLoS Comput Biol 9(8): e1003118. doi:10.1371/journal.pcbi.1003118 - Lionel Henry and Hadley Wickham (2020). purrr: Functional Programming Tools. R package version 03.4. https://CRAN.R-project.org/package=purrr - Love, M.I., Huber, W., Anders, S. Moderated estimation of fold change and dispersion for RNA-seq dta with DESeq2 Genome Biology 15(12):550 (2014) - Martin Morgan, Valerie Obenchain, Jim Hester and Hervé Pagès (2021). SummarizedExperiment: SmmarizedExperiment container. R package version 1.22.0. htps://bioconductor.org/packages/SummarizedExperiment - Orchestrating high-throughput genomic analysis with Bioconductor. W. Huber, V.J. Carey, R. Gntleman, ..., M. Morgan Nature Methods, 2015:12, 115. - Orchestrating high-throughput genomic analysis with Bioconductor. W. Huber, V.J. Carey, R. Gntleman, ..., M. Morgan Nature Methods, 2015:12, 115. - R Core Team (2021). R: A language and environment for statistical computing. R Foundation for Satistical Computing, Vienna, Austria. URL https://www.R-project.org/. - Sonali Arora, Martin Morgan, Marc Carlson and H. Pagès (2021). GenomeInfoDb: Utilities for mnipulating chromosome names, including modifying them to follow a particular naming style. R pckage version 1.28.0. https://bioconductor.org/packages/GenomeInfoDb - Wickham et al., (2019). Welcome to the tidyverse. Journal of Open Source Software, 4(43), 1686, https://doi.org/10.21105/joss.01686