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Kai Hu committed Jun 13, 2022
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70 changes: 70 additions & 0 deletions README.md
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- [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)
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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).
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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
PBMC_10K_N,/path/atac_pbmc_10k_nextgem_fastqs/atac_pbmc_10k_nextgem_S1_L002_R1_001.fastq.gz,/path/atac_pbmc_10k_nextgem_fastqs/atac_pbmc_10k_nextgem_S1_L002_R3_001.fastq.gz,/path/atac_pbmc_10k_nextgem_fastqs/atac_pbmc_10k_nextgem_S1_L002_R2_001.fastq.gz
PBMC_1K_N,/path/atac_pbmc_1k_nextgem_fastqs/atac_pbmc_1k_nextgem_S1_L001_R1_001.fastq.gz,/path/atac_pbmc_1k_nextgem_fastqs/atac_pbmc_1k_nextgem_S1_L001_R3_001.fastq.gz,/path/atac_pbmc_1k_nextgem_fastqs/atac_pbmc_1k_nextgem_S1_L001_R2_001.fastq.gz
PBMC_1K_N,/path/atac_pbmc_1k_nextgem_fastqs/atac_pbmc_1k_nextgem_S1_L002_R1_001.fastq.gz,/path/atac_pbmc_1k_nextgem_fastqs/atac_pbmc_1k_nextgem_S1_L002_R3_001.fastq.gz,/path/atac_pbmc_1k_nextgem_fastqs/atac_pbmc_1k_nextgem_S1_L002_R2_001.fastq.gz
PBMC_500_N,/path/atac_pbmc_500_nextgem_fastqs/atac_pbmc_500_nextgem_S1_L001_R1_001.fastq.gz,/path/atac_pbmc_500_nextgem_fastqs/atac_pbmc_500_nextgem_S1_L001_R3_001.fastq.gz,/path/atac_pbmc_500_nextgem_fastqs/atac_pbmc_500_nextgem_S1_L001_R2_001.fastq.gz
PBMC_500_N,/path/atac_pbmc_500_nextgem_fastqs/atac_pbmc_500_nextgem_S1_L002_R1_001.fastq.gz,/path/atac_pbmc_500_nextgem_fastqs/atac_pbmc_500_nextgem_S1_L002_R3_001.fastq.gz,/path/atac_pbmc_500_nextgem_fastqs/atac_pbmc_500_nextgem_S1_L002_R2_001.fastq.gz
PBMC_5K_N,/path/atac_pbmc_5k_nextgem_fastqs/atac_pbmc_5k_nextgem_S1_L001_R1_001.fastq.gz,/path/atac_pbmc_5k_nextgem_fastqs/atac_pbmc_5k_nextgem_S1_L001_R3_001.fastq.gz,/path/atac_pbmc_5k_nextgem_fastqs/atac_pbmc_5k_nextgem_S1_L001_R2_001.fastq.gz
PBMC_5K_N,/path/atac_pbmc_5k_nextgem_fastqs/atac_pbmc_5k_nextgem_S1_L002_R1_001.fastq.gz,/path/atac_pbmc_5k_nextgem_fastqs/atac_pbmc_5k_nextgem_S1_L002_R3_001.fastq.gz,/path/atac_pbmc_5k_nextgem_fastqs/atac_pbmc_5k_nextgem_S1_L002_R2_001.fastq.gz
PBMC_10K_V,/path/atac_v1_pbmc_10k_fastqs/atac_v1_pbmc_10k_S1_L001_R1_001.fastq.gz,/path/atac_v1_pbmc_10k_fastqs/atac_v1_pbmc_10k_S1_L001_R3_001.fastq.gz,/path/atac_v1_pbmc_10k_fastqs/atac_v1_pbmc_10k_S1_L001_R2_001.fastq.gz
PBMC_10K_V,/path/atac_v1_pbmc_10k_fastqs/atac_v1_pbmc_10k_S1_L002_R1_001.fastq.gz,/path/atac_v1_pbmc_10k_fastqs/atac_v1_pbmc_10k_S1_L002_R3_001.fastq.gz,/path/atac_v1_pbmc_10k_fastqs/atac_v1_pbmc_10k_S1_L002_R2_001.fastq.gz
PBMC_5K_V,/path/atac_v1_pbmc_5k_fastqs/atac_v1_pbmc_5k_S1_L001_R1_001.fastq.gz,/path/atac_v1_pbmc_5k_fastqs/atac_v1_pbmc_5k_S1_L001_R3_001.fastq.gz,/path/atac_v1_pbmc_5k_fastqs/atac_v1_pbmc_5k_S1_L001_R2_001.fastq.gz
PBMC_5K_V,/path/atac_v1_pbmc_5k_fastqs/atac_v1_pbmc_5k_S1_L002_R1_001.fastq.gz,/path/atac_v1_pbmc_5k_fastqs/atac_v1_pbmc_5k_S1_L002_R3_001.fastq.gz,/path/atac_v1_pbmc_5k_fastqs/atac_v1_pbmc_5k_S1_L002_R2_001.fastq.gz
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params {
profile = 'singularity,lsf'
}
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params {
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_blacklist = '/home/hl84w/lucio_castilla/scATAC-seq/docs/hg38-blacklist.v2.bed.gz'
archr_scrnaseq = '/path/scRNA-Hematopoiesis-Granja-2019.rds'
}
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