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Estimation motifs #78

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26 changes: 18 additions & 8 deletions README.md
Original file line number Diff line number Diff line change
Expand Up @@ -29,22 +29,28 @@ conda activate masterenv
4. Set the wd parameter in the nextflow.config file as path where the repository is saved. For example: '~/masterJLU2018/'.


**Important Note:** For conda the channel bioconda needs to be set as highest priority! This is required due to two different packages with the same name in different channels. For the pipeline the package jellyfish from the channel bioconda is needed and **NOT** the jellyfish package from the channel conda-forge!

**Important Notes:**
1. For conda the channel bioconda needs to be set as highest priority! This is required due to two different packages with the same name in different channels. For the pipeline the package jellyfish from the channel bioconda is needed and **NOT** the jellyfish package from the channel conda-forge!
2. The parameters --create_known_tfbs_path and --tfbs_path need an absolute path. If a relative path is given it will not work due to nextflow changing the working directory. This will soon be updated.


## Quick Start
```console
nextflow run pipeline.nf --bigwig [BigWig-file] --bed [BED-file] --genome_fasta [FASTA-file] --motif_db [MEME-file] --organism [mm10|mm9|hg19|hg38]
nextflow run pipeline.nf --bigwig [BigWig-file] --bed [BED-file] --genome_fasta [FASTA-file] --motif_db [MEME-file] --organism [mm10|mm9|hg19|hg38] (--tfbs_path [absolute PATH] || --create_known_tfbs_path [absolute PATH])
```

### Demo run
There are files provided inside ./demo/ for a demo run.
Go to the main directory and run following command:
```
nextflow run pipeline.nf --bigwig ./demo/buenrostro50k_chr1_fp.bw --bed ./demo/buenrostro50k_chr1_peaks.bed --genome_fasta ./demo/hg38/hg38_chr1.fa --motif_db ./demo/motif_database/jaspar_vertebrates.meme --out ./demo/buenrostro50k_chr1_out/ --create_known_tfbs_path ./demo/known_tfbs_hg38_chr1/ --organism hg38
nextflow run pipeline.nf --bigwig ./demo/buenrostro50k_chr1_fp.bw --bed ./demo/buenrostro50k_chr1_peaks.bed --genome_fasta ./demo/hg38/hg38_chr1.fa --motif_db ./demo/motif_database/jaspar_vertebrates.meme --out ./demo/buenrostro50k_chr1_out/ --create_known_tfbs_path [absolute path]/demo/known_tfbs/ --organism hg38
```
**Important Note:** It can happen that tfbsscan does not work due to an unknown bug. If this is the case you will see the error message shown in "Known Issues". If the error occurs first try to change the tfbsscan_method to fimo. If it persists try the following command:
```
nextflow run pipeline.nf --bigwig ./demo/buenrostro50k_chr1_fp.bw --bed ./demo/buenrostro50k_chr1_peaks.bed --genome_fasta ./demo/hg38/hg38_chr1.fa --motif_db ./demo/motif_database/jaspar_vertebrates.meme --out ./demo/buenrostro50k_chr1_out/ --tfbs_path [absolute path]/demo/known_tfbs_chr1_hg38/ --organism hg38
```


## Parameters
For a detailed overview for all parameters follow this [link](https://github.molgen.mpg.de/loosolab/masterJLU2018/wiki/Configuration).
```
Expand All @@ -56,14 +62,17 @@ Required arguments:
--config Path to UROPA configuration file
--organism Input organism [hg38 | hg19 | mm9 | mm10]
--out Output Directory (Default: './out/')
[
--create_known_tfbs_path Path to directory where output from tfbsscan (known motifs) are stored.
Path can be set as tfbs_path in next run. (Needs absolute path)
or
--tfbs_path Path to directory with output from tfbsscan. If given tfbsscan will be skipped. (Needs absolute path)
]

Optional arguments:

--help [0|1] 1 to show this help message. (Default: 0)
--tfbs_path Path to directory with output from tfbsscan. If given tfbsscan will not be run.
--create_known_tfbs_path Path to directory where output from tfbsscan (known motifs) are stored.
Path can be set as tfbs_path in next run. (Default: './')
--gtf_path Path to gtf-file. If path is set the process which creats a gtf-file is skipped.
--gtf_path Path to gtf-file. If path is set the process which creats a gtf-file is skipped.

Footprint extraction:
--window_length INT This parameter sets the length of a sliding window. (Default: 200)
Expand Down Expand Up @@ -97,6 +106,7 @@ Optional arguments:
--iteration INT Number of iterations done by GLAM2. More Iterations: better results, higher runtime. (Default: 10000)
--tomtom_treshold FLOAT Threshold for similarity score. (Default: 0.01)
--best_motif INT Get the best X motifs per cluster. (Default: 3)
--gap_penalty INT Set penalty for gaps in GLAM2 (Default: 1000)
Moitf clustering:
--cluster_motif Boolean If 1 pipeline clusters motifs. If its 0 it does not. (Default: 0)
--edge_weight INT Minimum weight of edges in motif-cluster-graph (Default: 5)
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4 changes: 2 additions & 2 deletions bin/2.2_motif_estimation/get_motif_seq.R
Original file line number Diff line number Diff line change
Expand Up @@ -51,12 +51,12 @@ create_seq_json <- function(input, output, num, tmp_path, cluster_id) {
stop(paste0("CLUSTER ID could not be found. Please make sure that your file path contains _[cluster_id] at the end. Found: ", cluster_id,"\n For example: /test_cluster_1/glam.txt"))
}

dir.create(tmp_path, showWarnings = FALSE)
dir.create(tmp_path, showWarnings = FALSE, recursive = TRUE)

file_dir <- tmp_path

# Split glam.txt file on lines that start with Score:
system(paste0("csplit ", input, " '/^Score:.*/' '{*}' -f ", file_dir, "/f_id_test.pholder"))
system(paste0("csplit ", input, " '/^Score:.*/' '{*}' -f ", file_dir, "/f_id.pholder"))
# Only keep the lines that start with 'f' to get the lines with the sequence ids
system(paste0("for i in ", file_dir, "/*.pholder0[1-", num, "];do grep \"^f\" $i > \"${i}.done\";done"))

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