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Important

We have now implemented the xf flag filtering in SnapATAC2-scooby:


subset-bam

subset-bam is a tool to subset a 10x Genomics BAM file based on a tag, most commonly the cell barcode tag. The latest version is v1.1.0 and can be found on the releases page.

Overview of how it works

subset-bam is a simple tool implemented in Rust that takes a 10x Genomics BAM file, a CSV file defining the subset of cells you want to isolate, and produces a new BAM file with only alignments associated with those cells.

In the subsetting process, temporary BAM files will be written to your temporary file ($TMPDIR) location before a final concatenation step. Please make sure this location is writeable has enough space to support this operation. If you cannot write to this location, you can set your TMPDIR variable to a path you can write by the command export TMPDIR=/my/writeable/path.

Support

This tool is not officially supported. If you have any comments, please submit a GitHub issue.

Installation

subset-bam has automatically generated downloadable binaries for generic linux and Mac OSX under the releases page. The linux binaries are expected to work on our supported Operating Systems.

Compiling from source

subset-bam is standard Rust executable project, that works with stable Rust >=1.13. Install Rust through the standard channels, then type cargo build --release. The executable will appear at target/release/subset-bam. As usual it's important to use a release build to get good performance.

Usage

--bam (-b): Input 10x Genomics BAM. This BAM must have the CB tag to define the barcodes of cell barcodes (or the tag defined by --bam-tag). Must also have an index (.bai) file. REQUIRED.

--cell-barcodes (-c): A cell barcodes file as produced by Cell Ranger that defines which barcodes were called as cells. One barcode per line. In Cell Ranger runs, this can be found in the sub-folder outs/filtered_gene_bc_matrices_mex/${refGenome}/barcodes.tsv where ${refGenome} is the name of the reference genome used in your Cell Ranger run. This file can be used as column labels for the output matrix. REQUIRED.

--out-bam (-o): A path to write the subsetted BAM file to. REQUIRED.

--cores: Number of parallel cores to use. DEFAULT: 1.

--log-level: One of info, error or debug. Increasing levels of logging. DEFAULT: error.

--bam-tag: Change this to use an alternative tag to the default CB tag. This can be useful for subsetting BAMs from LongRanger.

License

subset-bam is licensed under the MIT license. This project may not be copied, modified, or distributed except according to those terms.

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