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samples: samples.csv
ref:
species: "Drosophila melanogaster"
genome: ""
annotation: ""
accession: "GCF_000001215.4"
ensembl_species: "" # e.g., "homo_sapiens"
build: "" # e.g., "GRCh38"
release: "" # e.g., "105"
read_filter:
min_length: 200
minimap2:
index_opts: ""
opts: ""
maximum_secondary: 100
secondary_score_ratio: 1.0
samtools:
samtobam_opts: "-b"
bamsort_opts: ""
bamindex_opts: ""
bamstats_opts: ""
quant:
salmon_libtype: "U"
deseq2:
fit_type: ""
design_factors:
- "condition"
lfc_null: 1.0
alt_hypothesis: "greaterAbs"
point_width: 20
mincount: 10
alpha: 0.05
threshold_plot: 10
colormap: "Blues"
figtype: "png"
batch_effect:
- ""
isoform_analysis:
FLAIR: true
qscore: 1
exp_thresh: 10
col_opts: "--annotation_reliant generate --generate_map --stringent"
protein_annotation:
lambda: false
uniref: "https://ftp.imp.fu-berlin.de/pub/lambda/index/lambda3/gen_0/uniref50_20230713.lba.gz"
num_matches: 3
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This workflow performs differential expression analysis of RNA-seq data obtained from Oxford Nanopore long-read sequencing technology.
First a transcriptome FASTA is constructed using gffread. Reads are then mapped to the transcriptome with the long-read optimized alignment tool minimap2.
Next quantification is performed using salmon before normalization and differential expression analysis are conducted by PyDESeq2.
The workflow can optionally analyze splice-isoforms through integrating the FLAIR workflow.
Additionaly, NanoPlot is employed to analyze initial sequencing data and QualiMap is used to evaluate mapping results.
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