mumerge 0.0.8

A tool for combining bed regions from multiple bed files in a probabilistically prinipled manner.

A tool for combining bed regions from multiple bed files in a probabilistically-prinipled manner.

Installation

It is recommended to install mumerge using a virtual environment or package manager—e.g. venv or conda. Specifically, because bedtools must be available at the command line we recommend you create a new environment with conda and install bedtools from bioconda, as follows:

(base) $ conda create -n mumerge_env
(base) $ conda activate mumerge_env
(mumerge_env) $ conda install -c bioconda bedtools

To confirm installation, check the bedtools version:

(mumerge_env) $ bedtools --version
bedtools v2.30.0

Now, with bedtools available within your environment, you can install mumerge as follows.

Via pip

The simplest way of installing mumerge within your virtual environment is using pip. Be sure to use the appropriate version of Python if you have multiple versions installed. mumerge can then be installed with one of the following commands.

From PyPI (recommended):

$ python -m pip install mumerge

If successful, mumerge should now be callable from the command line.

From GitHub:

$ python -m pip install git+https://github.com/Dowell-Lab/mumerge

In order to upgrade to the latest version of mumerge from a previous one, include --upgrade in other of the previous pip commands.

Via git clone

Alternatively, you can download mumerge and all supporting files by cloning the GitHub repository to your local machine using git:

$ git clone https://github.com/Dowell-Lab/mumerge.git

If you clone the repo, you may want to add directory mumerge/mumerge to your system PATH variable (this will depend on your platform/OS) so that you can run mumerge directly from the command-line.

Dependencies

• Python>=3.6 https://www.python.org/downloads/

• NumPy https://numpy.org/

• bedtools https://bedtools.readthedocs.io/en/latest/content/installation.html

NumPy will be installed automatically when using pip to install mumerge. However, bedtools must be installed manually and made available in your system path prior to running mumerge.

Bedtools

muMerge relies on bedtools in order to group together those bed regions from the input bed files that will be combined probabilistically by muMerge. This grouping is done using the bedtools merge command, which must be available at the command line.

Usage

For general usage, see the help menu:

$ mumerge -h

This will return the general commands needed to run muMerge:

usage: mumerge.py [-h] [-H] [-i INPUT] [-o OUTPUT] [-w WIDTH] [-m MERGED] [-r] [-v]

Merges region calls (mu) generated by Tfit, or other peak calling functions across
multiple samples and replicates.

optional arguments:
  -h, --help            show this help message and exit
  -H, --HELP            Verbose help info about the input format.
  -i INPUT, --input INPUT
                        Input file (full path) containing bedfiles, sample ID's and
                        replicate grouping names (tab delimited). Each sample on separate
                        line. First line header, equal to '#file<TAB>sampid<TAB>group',
                        required. 'file' must be full path. 'sampid' can be any string.
                        'group' can be string or integer. See '-H' help flag for more
                        information.
  -o OUTPUT, --output OUTPUT
                        Output file basename (full path, sans extension). WARNING:
                        will overwrite any existing file)
  -w WIDTH, --width WIDTH
                        The ratio of a the sigma for the corresponding probabilty
                        distribution to the bed region (half-width) --- sigma:half-bed
                        (default: 1). The choice for this parameter will depend on the
                        data type as well as how bed regions were inferred from the
                        expression data.
  -m MERGED, --merged MERGED
                        Sorted bedfile (full path) containing the regions over which
                        to combine the sample bedfiles. If not specified, mumerge will
                        generate one directly from the sample bedfiles.
  -r, --remove_singletons
                        Remove calls not present in more than 1 sample
  -v, --verbose         Verbose printing during processing.

Input file

The <INPUT> file is a tab delimited text file that contains paths to BED files to be merged along with sample names as condition/replicate information for each sample. In the example below, there are 4 samples with two treatment groups.

#file   sampid  group
/path/to/sample1.bed    sample1 control
/path/to/sample2.bed    sample2 control
/path/to/sample3.bed    sample3 treatment
/path/to/sample4.bed    sample4 treatment

You can find this information using the -H flag—i.e. running mumerge -H, which will return the following:

INPUT FILE
----------
Input file containing bedfiles, sample ID's, and replicate groupings. Input
file (indicated by the '-i' flag) should be of the following (tab delimited)
format:

#file   sampid  group
/full/path/file1.bed    sampid1 A
/full/path/file2.bed    sampid2 B
...

Header line indicated by '#' character must be included and fields must
follow the same order as non-header lines. The order of subsequent lines does
not matter. File paths should be full paths to bed files, however you can
also specify paths that are relative to the input file location. 'group'
identifiers should group files that are technical/biological replicates.
Different experimental conditions should recieve different 'group' identifiers.
The 'group' identifier can be of type 'int' or 'str'. If 'sampid' is not
specified, then default sample ID's will be used.

Output files

muMerge returns the merged regions in BED file format (project_id_MUMERGE.bed). Additionally, a log file (project_id.log) that details the summary of the run is also inlcuded along with intermediate files (project_id_MISCALLS.bed and project_id_BEDTOOLS_MERGE.bed).

Demo

The additional help menu (mumerge -H) also contains information on a muMerge demo included with the package. The menu will specify where the demo files are located (install location depends on the platform) and how to run them. The demo consists of an input muMerge file which references two short bedfiles (a.bed and b.bed) that are located in the same directory. Running the demo (replace <fullpath> with the path to the input file which depends on where you installed it):

$ mumerge -v -i <fullpath>/mumerge_demo.input -o ./demo_out

will return the following to stdout:

Generating 'bedtools merge' bedfile...
Building bed-regions dictionary...
# Sample_ID      Filename
# sampA          <fullpath>/a.bed
# sampB          <fullpath>/b.bed
Processed 2 of 2 regions

and will produce the following files:

./demo_out.log
./demo_out_BEDTOOLS_MERGE.bed
./demo_out_MISCALLS.bed
./demo_out_MUMERGE.bed

If run correctly, demo_out_MUMERGE.bed should have two bed lines (chr1 150 350 and chr1 600 900), demo_out_MISCALLS.bed should be empty, and demo_out.log should contain meta information about the run.

Platforms

• Linux

• macOS

• Windows Subsystem for Linux (WSL)

Runtime

The overall run time depends on the the number of input BED files and regions being merged. A test case, where 8 samples (~30,000 regions) with 6 condition groups were merged, took about 12 minutes on a MacBook Pro iCore i9 2.3 GHz running macOS v 10.14.6.

Cite

Please cite the following article if you use muMerge: Transcription factor enrichment analysis (TFEA) quantifies the activity of multiple transcription factors from a single experiment

BibTeX citation:

@article{rubin2021transcription,
  title={Transcription factor enrichment analysis (TFEA) quantifies the activity of multiple transcription factors from a single experiment},
  author={Rubin, Jonathan D and Stanley, Jacob T and Sigauke, Rutendo F and Levandowski, Cecilia B and Maas, Zachary L and Westfall, Jessica and Taatjes, Dylan J and Dowell, Robin D},
  journal={Communications biology},
  volume={4},
  number={1},
  pages={1--15},
  year={2021},
  publisher={Nature Publishing Group}
}