nf-core · vagkaratzas · Dec 2, 2025 · Dec 1, 2025 · Dec 1, 2025 · Dec 2, 2025
diff --git a/CHANGELOG.md b/CHANGELOG.md
@@ -9,13 +9,15 @@ Initial release of nf-core/proteinannotator, created with the [nf-core](https://
 
 ### `Added`
 
-- [[PR #52](https://github.com/nf-core/proteinannotator/pull/52)] Add option to turn off InterProScan for testing
-- [[PR #51](https://github.com/nf-core/proteinannotator/pull/51)] Update to nf-core/tools v3.3.1
-- [[PR #47](https://github.com/nf-core/proteinannotator/pull/47)] Update metromap with more tools added from [May 2025 Hackathon](https://nf-co.re/events/2025/hackathon-boston)
-- [[PR #43](https://github.com/nf-core/proteinannotator/pull/44)] Add [mTM-Align](https://nf-co.re/modules/mtmalign_align/) and [MMseqs2 Search](https://nf-co.re/modules/mmseqs_search/) modules
-- [[PR #42](https://github.com/nf-core/proteinannotator/pull/42)] Updated to `nf-test` on GitHub Actions and in the `PULL_REQUEST_TEMPLATE.md`
-- [[PR #13](https://github.com/nf-core/proteinannotator/pull/13)] Add nf-core seqkit/stats module
-- [[PR #9](https://github.com/nf-core/proteinannotator/pull/9)] Add [InterProScan](https://interproscan-docs.readthedocs.io/) module
+- [#59](https://github.com/nf-core/proteinannotator/pull/59) - Added nf-core qc and pre-processing subworkflow for amino acid sequences `FAA_SEQFU_SEQKIT`. (by @vagkaratzas). (by @vagkaratzas)
+- [#57](https://github.com/nf-core/proteinannotator/pull/57) - nf-core tools template update to 3.5.1. (by @vagkaratzas)
+- [#52](https://github.com/nf-core/proteinannotator/pull/52) - Add option to turn off InterProScan for testing
+- [#51](https://github.com/nf-core/proteinannotator/pull/51) - Update to nf-core/tools v3.3.1
+- [#47](https://github.com/nf-core/proteinannotator/pull/47) - Update metromap with more tools added from [May 2025 Hackathon](https://nf-co.re/events/2025/hackathon-boston)
+<!-- - [#43](https://github.com/nf-core/proteinannotator/pull/44) - Add [mTM-Align](https://nf-co.re/modules/mtmalign_align/) and [MMseqs2 Search](https://nf-co.re/modules/mmseqs_search/) modules -->
+- [#42](https://github.com/nf-core/proteinannotator/pull/42) - Updated to `nf-test` on GitHub Actions and in the `PULL_REQUEST_TEMPLATE.md`
+- [#13](https://github.com/nf-core/proteinannotator/pull/13) - Add nf-core seqkit/stats module
+- [#9](https://github.com/nf-core/proteinannotator/pull/9) - Add [InterProScan](https://interproscan-docs.readthedocs.io/) module
 
 ### `Fixed`
 

diff --git a/CITATIONS.md b/CITATIONS.md
@@ -10,21 +10,29 @@
 
 ## Pipeline tools
 
-- [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/)
+- [SeqFu](https://pubmed.ncbi.nlm.nih.gov/34066939/)
 
-> Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924.
+> Telatin A, Fariselli P, Birolo G. SeqFu: a suite of utilities for the robust and reproducible manipulation of sequence files. Bioengineering. 2021 May 7;8(5):59. doi: 10.3390/bioengineering8050059. PubMed PMID: 34066939; PubMed Central PMCID: PMC8148589.
+
+- [SeqKit](https://pubmed.ncbi.nlm.nih.gov/38898985/)
+
+> Shen W, Sipos B, Zhao L. SeqKit2: A Swiss army knife for sequence and alignment processing. iMeta. 2024 Apr 5:e191. doi: 10.1002/imt2.191. PubMed PMID: 38898985; PubMed Central PMCID: PMC11183193.
 
 - [InterProScan](https://academic.oup.com/bioinformatics/article/17/9/847/206564)
 
 > Zdobnov, Evgeni M., and Rolf Apweiler. “InterProScan – an Integration Platform for the Signature-Recognition Methods in InterPro.” Bioinformatics 17, no. 9 (September 1, 2001): 847–48. https://doi.org/10.1093/bioinformatics/17.9.847.
 
-- [MMseqs2](https://www.nature.com/articles/nbt.3988)
+<!-- - [MMseqs2](https://www.nature.com/articles/nbt.3988)
 
 > Zdobnov, Evgeni M., and Rolf Apweiler. “InterProScan – an Integration Platform for the Signature-Recognition Methods in InterPro.” Bioinformatics 17, no. 9 (September 1, 2001): 847–48. https://doi.org/10.1093/bioinformatics/17.9.847.
 
 - [mTM-align](https://academic.oup.com/bioinformatics/article/34/10/1719/4769500)
 
-> Dong, Runze, Zhenling Peng, Yang Zhang, and Jianyi Yang. “mTM-Align: An Algorithm for Fast and Accurate Multiple Protein Structure Alignment.” Bioinformatics 34, no. 10 (May 15, 2018): 1719–25. https://doi.org/10.1093/bioinformatics/btx828.
+> Dong, Runze, Zhenling Peng, Yang Zhang, and Jianyi Yang. “mTM-Align: An Algorithm for Fast and Accurate Multiple Protein Structure Alignment.” Bioinformatics 34, no. 10 (May 15, 2018): 1719–25. https://doi.org/10.1093/bioinformatics/btx828. -->
+
+- [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/)
+
+> Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924.
 
 ## Software packaging/containerisation tools
 

diff --git a/assets/methods_description_template.yml b/assets/methods_description_template.yml
@@ -3,7 +3,6 @@ description: "Suggested text and references to use when describing pipeline usag
 section_name: "nf-core/proteinannotator Methods Description"
 section_href: "https://github.com/nf-core/proteinannotator"
 plot_type: "html"
-## TODO nf-core: Update the HTML below to your preferred methods description, e.g. add publication citation for this pipeline
 ## You inject any metadata in the Nextflow '${workflow}' object
 data: |
   <h4>Methods</h4>

diff --git a/conf/modules.config b/conf/modules.config
@@ -18,6 +18,62 @@ process {
         saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
     ]
 
+    withName: 'NFCORE_PROTEINANNOTATOR:PROTEINANNOTATOR:FAA_SEQFU_SEQKIT:SEQFU_STATS_BEFORE' {
+        ext.prefix = { "${meta.id}_before" }
+        publishDir = [
+            path: { "${params.outdir}/qc/${meta.id}/" },
+            mode: params.publish_dir_mode,
+            saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
+        ]
+    }
+
+    withName: 'NFCORE_PROTEINANNOTATOR:PROTEINANNOTATOR:FAA_SEQFU_SEQKIT:SEQFU_STATS_AFTER' {
+        ext.prefix = { "${meta.id}_after" }
+        publishDir = [
+            path: { "${params.outdir}/qc/${meta.id}/" },
+            mode: params.publish_dir_mode,
+            saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
+        ]
+    }
+
+    withName: 'NFCORE_PROTEINANNOTATOR:PROTEINANNOTATOR:FAA_SEQFU_SEQKIT:SEQKIT_SEQ' {
+        ext.args = [
+            "--remove-gaps",
+            "--upper-case",
+            "--validate-seq",
+            "--min-len ${params.min_seq_length}",
+            "--max-len ${params.max_seq_length}"
+        ].join(' ').trim()
+        ext.prefix = "intermediate_seqkit_seq"
+        publishDir = [
+            path: { "${params.outdir}/qc/${meta.id}/" },
+            mode: params.publish_dir_mode,
+            enabled: false,
+            saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
+        ]
+    }
+
+    withName: 'NFCORE_PROTEINANNOTATOR:PROTEINANNOTATOR:FAA_SEQFU_SEQKIT:SEQKIT_REPLACE' {
+        ext.args   = '-p "/" -r "_"'
+        ext.suffix = "fasta"
+        ext.prefix = "intermediate_seqkit_replace"
+        publishDir = [
+            path: { "${params.outdir}/qc/${meta.id}/" },
+            mode: params.publish_dir_mode,
+            enabled: false,
+            saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
+        ]
+    }
+
+    withName: 'NFCORE_PROTEINANNOTATOR:PROTEINANNOTATOR:FAA_SEQFU_SEQKIT:SEQKIT_RMDUP' {
+        ext.args   = { params.remove_duplicates_on_sequence ? "--by-seq" : '' }
+        publishDir = [
+            path: { "${params.outdir}/qc/${meta.id}/" },
+            mode: params.publish_dir_mode,
+            saveAs: { filename -> filename.equals('versions.yml') ? null : filename }
+        ]
+    }
+
     withName: 'MULTIQC' {
         ext.args   = { params.multiqc_title ? "--title \"$params.multiqc_title\"" : '' }
         publishDir = [
@@ -27,7 +83,4 @@ process {
         ]
     }
 
-    withName: SEQKIT_STATS {
-        ext.args = ' ' // turn off --all default argument
-    }
 }
diff --git a/docs/output.md b/docs/output.md
@@ -12,12 +12,51 @@ The directories listed below will be created in the results directory after the
 
 The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps:
 
+- [Quality check and preprocessing](#quality-check-and-preprocessing)
+  - [SeqFu](#seqfu) for input amino acid sequences quality check (QC)
+  - [SeqKit](#seqkit) for preprocessing input amino acid sequences (i.e., gap removal, convert to upper case, validate, filter by length, replace special characters such as `/`, and remove duplicate sequences)
+
 - [Functional Annotation](#functional-annotation) Annotate proteins with functional domains
   - [InterProScan](#Interproscan) - Search the InterPro database for functional domains
 - [MultiQC](#multiqc) - Aggregate report describing results and QC from the whole pipeline
-- [SeqKit stats](#seqkit_stats) - Simple statistics for protein FASTA files
 - [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution
 
+### Quality check and preprocessing
+
+#### SeqFu
+
+<details markdown="1">
+<summary>Output files</summary>
+
+- `qc/`
+  - `<samplename>/`
+    - `<samplename>_before.tsv`: Statistics for the input amino acid sequences before preprocessing
+    - `<samplename>_before_mqc.txt`: Statistics for the input amino acid sequences in MultiQC-ready format before preprocessing
+    - `<samplename>_after.tsv`: (optional) Statistics for the input amino acid sequences after preprocessing
+    - `<samplename>_after_mqc.txt`: (optional) Statistics for the input amino acid sequences in MultiQC-ready format after preprocessing
+    - `<samplename>.log`: (optional) Output file with count of duplicate sequences that were found and removed
+
+</details>
+
+The `seqfu` module is used for statistics generation of input amino acid sequences, both before and after preprocessing.
+
+[SeqFu](https://github.com/telatin/seqfu2) is a cross-platform compiled suite of tools to manipulate and inspect `FASTA` and `FASTQ` files.
+
+#### SeqKit
+
+<details markdown="1">
+<summary>Output files</summary>
+
+- `qc/`
+  - `<samplename>/`
+    - `<samplename>.<suffix>`: Updated preprocessed input fasta file
+
+</details>
+
+The `seqkit` module is used for initial preprocessing (i.e., gap removal, convert to upper case, validate, filter by length, replace special characters such as `/`, and remove duplicate sequences) of the input amino acid sequences.
+
+[SeqKit](https://github.com/shenwei356/seqkit) is a cross-platform and ultrafast toolkit for FASTA/Q file manipulation.
+
 ### Functional Annotation
 
 #### InterProScan

diff --git a/main.nf b/main.nf
@@ -38,7 +38,8 @@ workflow NFCORE_PROTEINANNOTATOR {
     // WORKFLOW: Run pipeline
     //
     PROTEINANNOTATOR (
-        samplesheet
+        samplesheet,
+        params.skip_preprocessing
     )
     emit:
     multiqc_report = PROTEINANNOTATOR.out.multiqc_report // channel: /path/to/multiqc_report.html

diff --git a/modules.json b/modules.json
@@ -20,6 +20,26 @@
                         "git_sha": "af27af1be706e6a2bb8fe454175b0cdf77f47b49",
                         "installed_by": ["modules"]
                     },
+                    "seqfu/stats": {
+                        "branch": "master",
+                        "git_sha": "e753770db613ce014b3c4bc94f6cba443427b726",
+                        "installed_by": ["faa_seqfu_seqkit"]
+                    },
+                    "seqkit/replace": {
+                        "branch": "master",
+                        "git_sha": "41dfa3f7c0ffabb96a6a813fe321c6d1cc5b6e46",
+                        "installed_by": ["faa_seqfu_seqkit"]
+                    },
+                    "seqkit/rmdup": {
+                        "branch": "master",
+                        "git_sha": "41dfa3f7c0ffabb96a6a813fe321c6d1cc5b6e46",
+                        "installed_by": ["faa_seqfu_seqkit"]
+                    },
+                    "seqkit/seq": {
+                        "branch": "master",
+                        "git_sha": "41dfa3f7c0ffabb96a6a813fe321c6d1cc5b6e46",
+                        "installed_by": ["faa_seqfu_seqkit"]
+                    },
                     "seqkit/stats": {
                         "branch": "master",
                         "git_sha": "81880787133db07d9b4c1febd152c090eb8325dc",
@@ -34,6 +54,11 @@
             },
             "subworkflows": {
                 "nf-core": {
+                    "faa_seqfu_seqkit": {
+                        "branch": "master",
+                        "git_sha": "15c0a7968179d3b717a9973a1c4f25beb8a9aa2b",
+                        "installed_by": ["subworkflows"]
+                    },
                     "utils_nextflow_pipeline": {
                         "branch": "master",
                         "git_sha": "05954dab2ff481bcb999f24455da29a5828af08d",

diff --git a/modules/nf-core/seqfu/stats/environment.yml b/modules/nf-core/seqfu/stats/environment.yml
diff --git a/modules/nf-core/seqfu/stats/main.nf b/modules/nf-core/seqfu/stats/main.nf
diff --git a/modules/nf-core/seqfu/stats/meta.yml b/modules/nf-core/seqfu/stats/meta.yml