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Normalized/Raw data mishap on the report #271

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rekren opened this issue May 21, 2024 · 1 comment
Closed

Normalized/Raw data mishap on the report #271

rekren opened this issue May 21, 2024 · 1 comment
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bug Something isn't working

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@rekren
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rekren commented May 21, 2024

Description of the bug

Hi there,
I am using nf-core/differentialabundance v.1.5.0.

It seems that in the "Exploratory analysis" section of the "Results" part, Raw values and Normalized values needs to replace each other.
It looks like they are switched by mistake of the generated Rmarkdown code.

here are the representative plots of the generated -suspiciously- looking plots.
image
image

Command used and terminal output

nextflow run nf-core/differentialabundance -r 1.5.0 --input dif_ab_samplesheet.csv --matrix outs/merged_gene_counts_table.tsv --transcript_length_matrix outs/merged_gene_lengths_table.tsv --contrasts dif_ab_contrast.csv --gtf /home/rekren/save/refs/Homo_sapiens.GRCh38.111.gtf.gz -profile genotoul --outdir DiffAbundance --report_author 'Rüçhan Ekren' --logo_file inf_logo_smaller.png --report_title 'Report' --report_description ' Explanation of the report' --shinyngs_build_app --report_scree

Relevant files

{
"input": "dif_ab_samplesheet.csv",
"study_name": "study",
"study_type": "rnaseq",
"study_abundance_type": "counts",
"contrasts": "dif_ab_contrast.csv",
"querygse": null,
"matrix": "outs/merged_gene_counts_table.tsv",
"transcript_length_matrix": "outs/merged_gene_lengths_table.tsv",
"control_features": null,
"sizefactors_from_controls": false,
"logo_file": "inf_logo_smaller.png",
"css_file": "/home/rekren/.nextflow/assets/nf-core/differentialabundance/assets/nf-core_style.css",
"citations_file": "/home/rekren/.nextflow/assets/nf-core/differentialabundance/CITATIONS.md",
"report_file": "/home/rekren/.nextflow/assets/nf-core/differentialabundance/assets/differentialabundance_report.Rmd",
"report_title": Title",
"report_author": "R\u00fc\u00e7han Ekren",
"report_contributors": null,
"report_description": "This is description.",
"report_scree": true,
"report_round_digits": 4,
"observations_type": "sample",
"observations_id_col": "sample",
"observations_name_col": null,
"features": null,
"features_type": "gene",
"features_id_col": "gene_id",
"features_name_col": "gene_name",
"features_metadata_cols": "gene_id,gene_name,gene_biotype",
"features_gtf_feature_type": "transcript",
"features_gtf_table_first_field": "gene_id",
"affy_cel_files_archive": null,
"affy_file_name_col": "file",
"affy_background": true,
"affy_bgversion": 2,
"affy_destructive": false,
"affy_cdfname": null,
"affy_rm_mask": false,
"affy_rm_outliers": false,
"affy_rm_extra": false,
"affy_build_annotation": true,
"proteus_measurecol_prefix": "LFQ intensity",
"proteus_norm_function": "normalizeMedian",
"proteus_plotsd_method": "violin",
"proteus_plotmv_loess": true,
"proteus_palette_name": "Set1",
"filtering_min_samples": 1.0,
"filtering_min_abundance": 1.0,
"filtering_min_proportion": null,
"filtering_grouping_var": null,
"filtering_min_proportion_not_na": 0.5,
"filtering_min_samples_not_na": null,
"exploratory_main_variable": "auto_pca",
"exploratory_clustering_method": "ward.D2",
"exploratory_cor_method": "spearman",
"exploratory_n_features": 500,
"exploratory_whisker_distance": 1.5,
"exploratory_mad_threshold": -5,
"exploratory_assay_names": "raw,normalised,variance_stabilised",
"exploratory_final_assay": "variance_stabilised",
"exploratory_log2_assays": "raw,normalised",
"exploratory_palette_name": "Set1",
"differential_file_suffix": null,
"differential_feature_id_column": "gene_id",
"differential_feature_name_column": "gene_name",
"differential_fc_column": "log2FoldChange",
"differential_pval_column": "pvalue",
"differential_qval_column": "padj",
"differential_min_fold_change": 2.0,
"differential_max_pval": 1.0,
"differential_max_qval": 0.05,
"differential_foldchanges_logged": true,
"differential_palette_name": "Set1",
"differential_subset_to_contrast_samples": false,
"deseq2_test": "Wald",
"deseq2_fit_type": "parametric",
"deseq2_sf_type": "ratio",
"deseq2_min_replicates_for_replace": 7,
"deseq2_use_t": false,
"deseq2_lfc_threshold": 0,
"deseq2_alt_hypothesis": "greaterAbs",
"deseq2_independent_filtering": true,
"deseq2_p_adjust_method": "BH",
"deseq2_alpha": 0.1,
"deseq2_minmu": 0.5,
"deseq2_vs_method": "vst",
"deseq2_shrink_lfc": true,
"deseq2_cores": 1,
"deseq2_vs_blind": true,
"deseq2_vst_nsub": 1000,
"limma_ndups": null,
"limma_spacing": null,
"limma_block": null,
"limma_correlation": null,
"limma_method": "ls",
"limma_proportion": 0.01,
"limma_stdev_coef_lim": "0.1,4",
"limma_trend": false,
"limma_robust": false,
"limma_winsor_tail_p": "0.05,0.1",
"limma_adjust_method": "BH",
"limma_p_value": 1.0,
"limma_lfc": 0,
"limma_confint": false,
"gsea_run": false,
"gsea_nperm": 1000,
"gsea_permute": "phenotype",
"gsea_scoring_scheme": "weighted",
"gsea_metric": "Signal2Noise",
"gsea_sort": "real",
"gsea_order": "descending",
"gsea_set_max": 500,
"gsea_set_min": 15,
"gsea_norm": "meandiv",
"gsea_rnd_type": "no_balance",
"gsea_make_sets": true,
"gsea_median": false,
"gsea_num": 100,
"gsea_plot_top_x": 20,
"gsea_rnd_seed": "timestamp",
"gsea_save_rnd_lists": false,
"gsea_zip_report": false,
"gprofiler2_run": false,
"gprofiler2_organism": null,
"gprofiler2_significant": true,
"gprofiler2_measure_underrepresentation": false,
"gprofiler2_correction_method": "gSCS",
"gprofiler2_sources": null,
"gprofiler2_evcodes": false,
"gprofiler2_max_qval": 0.05,
"gprofiler2_token": null,
"gprofiler2_background_file": "auto",
"gprofiler2_background_column": null,
"gprofiler2_domain_scope": "annotated",
"gprofiler2_min_diff": 1,
"gprofiler2_palette_name": "Blues",
"shinyngs_build_app": true,
"shinyngs_guess_unlog_matrices": true,
"shinyngs_deploy_to_shinyapps_io": false,
"shinyngs_shinyapps_account": null,
"shinyngs_shinyapps_app_name": null,
"gene_sets_files": null,
"genome": null,
"igenomes_base": "s3://ngi-igenomes/igenomes/",
"igenomes_ignore": true,
"outdir": "DiffAbundance",
"publish_dir_mode": "copy",
"email": null,
"email_on_fail": null,
"plaintext_email": false,
"monochrome_logs": false,
"hook_url": null,
"help": false,
"version": false,
"config_profile_name": null,
"config_profile_description": "The Genotoul cluster profile",
"custom_config_version": "master",
"gtf": "/home/rekren/save/refs/Homo_sapiens.GRCh38.111.gtf.gz",
"custom_config_base": "https://raw.githubusercontent.com/nf-core/configs/master",
"config_profile_contact": "support.bioinfo.genotoul@inra.fr",
"config_profile_url": "http://bioinfo.genotoul.fr/",
"max_memory": {
"mega": 122880,
"kilo": 125829120,
"giga": 120,
"bytes": 128849018880
},
.nextflow.log

"max_cpus": 48,
"max_time": {
    "days": 4,
    "millis": 345600000,
    "hours": 96,
    "minutes": 5760,
    "seconds": 345600,
    "durationInMillis": 345600000
},
"validationFailUnrecognisedParams": false,
"validation-fail-unrecognised-params": false,
"validationLenientMode": false,
"validation-lenient-mode": false,
"validationSchemaIgnoreParams": "genomes,igenomes_base",
"validation-schema-ignore-params": "genomes,igenomes_base",
"validationShowHiddenParams": false,
"validation-show-hidden-params": false,
"validate_params": true,
"save_reference": true,
"igenomesIgnore": true,
"igenomes-ignore": true,
"genomes": {
    
}

}

System information

Nextflow version : version 23.10.0, build 5889
Hardware: HPC (Genotoul)
Executor: Slurm
Container: Singularity
OS: Linux
Version of nf-core/differentialabundance : 1.5

@rekren rekren added the bug Something isn't working label May 21, 2024
@rekren
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rekren commented May 21, 2024

@pinin4fjords and Oskar Wacker checked the label assignments with the test data on test profile, everything seemed good.

I have tested with another dataset and also tested with nf-core/differential_abundance v1.4.
They were seeming appropriately assigning the assay names.
I think everything is indeed in order and this was only visual "mishap".
For the above figures when I check the actual png figures they were looking a bit better than the rendered plots of the report.
image
image

For this specific dataset, I believe data plots shrunken a little bit more than usual and made us question the correctness of the labels on the report rendering part .

Closing the issue with this comments.

@rekren rekren closed this as completed May 21, 2024
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