From 216f1460577926a04a2d2918f36588afb6217f8a Mon Sep 17 00:00:00 2001 From: Kai Hu <> Date: Mon, 13 Jun 2022 13:46:18 -0400 Subject: [PATCH] minor --- README.md | 70 ++++++++++++++++++++++++++++++ assets/10X_human_scatac_fastq.csv | 13 ++++++ assets/hg38-blacklist.v2.bed.gz | Bin 0 -> 5867 bytes conf/tem.config | 3 ++ conf/test_chromap_initial.config | 10 +++++ docs/template_of_method.docx | Bin 32281 -> 31727 bytes 6 files changed, 96 insertions(+) create mode 100644 assets/10X_human_scatac_fastq.csv create mode 100644 assets/hg38-blacklist.v2.bed.gz create mode 100644 conf/tem.config create mode 100644 conf/test_chromap_initial.config diff --git a/README.md b/README.md index 72ff5ef..b7f6e76 100644 --- a/README.md +++ b/README.md @@ -9,6 +9,7 @@ - [DOWNSTREAM_ARCHR](#downstream_archr) [Quick Start](#quick-start) +[Example: plots in paper](#Example:-plots-in-paper) [Documentation](#documentation) [Credits](#credits) [Bug report/Support](#bug-reportsupport) @@ -143,6 +144,75 @@ nextflow run main.nf --help See documentation [usage](https://github.com/hukai916/scATACpipe/blob/main/docs/usage.md) for all of the available options. +## Example: plots in paper + +This section describes how the plots in the manuscript (to be added) were generated using scATACpipe. For comparison, the manuscript conducted 3 separate analyses, each using a different preprocessing strategy (`default`, `10xgenomics`, `chromap`). Since the commands and preprocessed results are quite similar across the three methods, only the `chromap` option will be demonstrated here. + +1. The initial execution: + +``` +nextflow run main.nf -profile singularity,lsf -c conf/test_chromap.config --preprocess chromap --outdir ./chromap_results_haibo --input_fastq /home/hl84w/lucio_castilla/scATAC-seq/docs/10X_human_scatac_fastq_new_style.csv --ref_fasta_ensembl homo_sapiens --species_latin_name 'homo sapiens' --archr_blacklist /home/hl84w/lucio_castilla/scATAC-seq/docs/hg38-blacklist.v2.bed.gz --doublet_removal_algorithm archr --archr_thread 8 -resume e209b4a2-1ada-4893-af13-132d1e3f5a55 +``` + +``` +nextflow run main.nf -profile singularity,lsf --preprocess chromap --outdir ./results_chromap --input_fastq ./assets/10X_human_scatac_fastq.csv --ref_fasta_ensembl homo_sapiens --species_latin_name 'homo sapiens' --archr_scrnaseq '/path/scRNA-Hematopoiesis-Granja-2019.rds' --archr_blacklist /home/hl84w/lucio_castilla/scATAC-seq/docs/hg38-blacklist.v2.bed.gz +``` + +Break down: + + * `-profile singularity,lsf`: + + This option instructs scATACpipe to use Singularity containers and LSF as the executor. Multiple parameters are separated by commas. Since `profile` is pipeline-level flag, it is prefixed with a single dash (`-`). Module-level flags are prefixed with double dash (`--`). + + * `--preprocess chromap`: + + This instructs scATACpipe to use Chromap preprocessing strategy. + + * `--outdir ./results_chromap`: + + Output will be saved into `./results_chromap` folder. + + * `--input_fastq ./assets/10X_human_scatac_fastq.csv`: + + Please replace the `/path/` in the `10X_human_scatac_fastq.csv` with absolute paths. Details regarding the 6 samples can be found in the supplementary section of the paper. If you detect any outlier samples, you can remove them from the downstream analyses using the `--filter_sample = 'PBMC_10K_C, PBMC_10K_X'` flag. + + * `--ref_fasta_ensembl homo_sapiens`: + + This specifies that the genome _Homo Sapiens_ from ENSEMBLE will be used as reference. To view all supported genomes, check out `nextflow run main.nf --support_genome`. + + * `--species_latin_name 'homo sapiens'`: + + Simply the Latin name of the reference genome. + + * `--archr_scrnaseq '/path/scRNA-Hematopoiesis-Granja-2019.rds'` + + Matching scRNA-seq data. Can ignore if not available. The example file can be downloaded [here](https://jeffgranja.s3.amazonaws.com/ArchR/TestData/scRNA-Hematopoiesis-Granja-2019.rds). + + * `--archr_blacklist ./assets/hg38-blacklist.v2.bed.gz`: + + Blacklist to exclude for downstream analysis. Click [here](https://github.com/Boyle-Lab/Blacklist/tree/master/lists) for other species. + +Instead of passing each flag option via the command line, you can include them all in a configuration file and supply it with the `-c` option. Below is equivalent to above: +``` +nextflow run main.nf -profile singularity,lsf -c ./conf/test_chromap_initial.config +``` + +2. The final execution: +After examining the results from the initial execution, we decided to remove the + + + --archr_thread 8 + + + + + + + + +To replicate the plots showed in the manuscript (to be added), + + ## Documentation The scATACpipe workflow comes with documentation about the pipeline: [usage](https://github.com/hukai916/scATACpipe/blob/main/docs/usage.md) and [output](https://github.com/hukai916/scATACpipe/blob/main/docs/output.md). diff --git a/assets/10X_human_scatac_fastq.csv b/assets/10X_human_scatac_fastq.csv new file mode 100644 index 0000000..c1d1231 --- /dev/null +++ b/assets/10X_human_scatac_fastq.csv @@ -0,0 +1,13 @@ +sample_name,path_fastq_1,path_fastq_2,path_barcode +PBMC_10K_N,/path/atac_pbmc_10k_nextgem_fastqs/atac_pbmc_10k_nextgem_S1_L001_R1_001.fastq.gz,/path/atac_pbmc_10k_nextgem_fastqs/atac_pbmc_10k_nextgem_S1_L001_R3_001.fastq.gz,/path/atac_pbmc_10k_nextgem_fastqs/atac_pbmc_10k_nextgem_S1_L001_R2_001.fastq.gz 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