Mercurial > repos > youngkim > ezbamqc
diff README.rst @ 6:773b3a97d43c
Uploaded
author | cshl-bsr |
---|---|
date | Tue, 29 Mar 2016 16:51:06 -0400 |
parents | |
children |
line wrap: on
line diff
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/README.rst Tue Mar 29 16:51:06 2016 -0400 @@ -0,0 +1,186 @@ +======= +ezBAMQC +======= + +*"ezBAMQC, a tool to check the quality of mapped next generation sequencing files."* + +:Codeology Icon: + + .. image:: https://raw.githubusercontent.com/mhammell-laboratory/bamqc/master/doc/bamqc-icon.gif + :alt: generated at codeology.braintreepayments.com/mhammell-laboratory/bamqc + :align: right + :target: http://codeology.braintreepayments.com/mhammell-laboratory/bamqc + +:Description: + + ezBAMQC is a tool to check the quality of either one or many mapped next-generation-sequencing datasets. It conducts comprehensive evaluations of aligned sequencing data from multiple aspects including: clipping profile, mapping quality distribution, mapped read length distribution, genomic/transcriptomic mapping distribution, inner distance distribution (for paired-end reads), ribosomal RNA contamination, transcript 5’ and 3’ end bias, transcription dropout rate, sample correlations, sample reproducibility, sample variations. It outputs a set of tables and plots and one HTML page that contains a summary of the results. Many metrics are designed for RNA-seq data specifically, but ezBAMQC can be applied to any mapped sequencing dataset such as RNA-seq, CLIP-seq, GRO-seq, ChIP-seq, DNA-seq and so on. + +:Links: + + `Github Page <https://github.com/mhammell-laboratory/bamqc>`_ + + `Pypi Page <https://pypi.python.org/pypi/ezBAMQC>`_ + + `MHammell Lab <http://hammelllab.labsites.cshl.edu/software>`_ + +:Authors: + Ying Jin, David Molik, and Molly Hammell + +:Version: 0.6.7 + +:Contact: + Ying Jin (yjin@cshl.edu) + +Installation guide for ezBAMQC for from source installs +======================================================= + +When installing ezBAMQC there are several options, but the main point is: since ezBAMQC uses C++ STD 11 you'll need a version of GCC that can support that, this useally means 4.8 or 4.9. beyond that, you'll need Python, R and Corrplot for interfacing with the C code. + +:Intallation: + `Source Code <https://github.com/mhammell-laboratory/ezBAMQC/releases>`_ + + `Pypi <https://pypi.python.org/pypi?:action=display&name=ezBAMQC>`_ + +:Prerequisites: + * `python2.7 <https://www.python.org/download/releases/2.7/>`_ + * `R <https://www.r-project.org/>`_ + * `corrplot <https://cran.r-project.org/web/packages/corrplot/>`_ + * `GCC 4.8.1 or greater <https://gcc.gnu.org/gcc-4.8/>`_ GCC 4.9.1 or greater is recomended for PyPi install + +:Notes: + * While there are multiple methods of installing the prerequistes it may help to look at (if using a yum based linux distro):* + * `Devtoolset-3 <https://access.redhat.com/documentation/en-US/Red_Hat_Developer_Toolset/3/html/User_Guide/sect-Red_Hat_Developer_Toolset-Install.html>`_ for GCC compilers + * `IUS <https://ius.io/>`_ for Python2.7 + * `Software Collections <https://www.softwarecollections.org/>`_ for collections of software (like devtoolset 3 or python) + * `rpmfinder <https://www.rpmfind.net/>`_ for searching rpms across mutliple systems + +Setup +===== + +1) Make sure that the GCC comiler is in your PATH: + +:: + + export PATH=/path/to/gcc:$PATH + +2) Make sure that python2.7 is in your PYTHONPATH: + +:: + + export PYTHONPATH=/path/to/python2.7/site-packages:$PYTHONPATH + +3) There are three methods of installation of ezBAMQC, from source, from setup.py, and from pypi, once prequistes are setup. + +From Source +~~~~~~~~~~~ + +1) Download source + +2) Unpack tarball and go to the directory of the package: + +:: + + tar xvfz bamqc-0.6.7.tar.gz + + cd bamqc-0.6.7 + +3) Run make: + +:: + + make + +From Setup.py +~~~~~~~~~~~~~ + +:: + + python2.7 setup.py install + +From Pypi +~~~~~~~~~ + +:: + + pip2.7 install BAMqc + +Usage +===== + +:: + + ezBAMQC [-h] -i alignment_files [alignment_files ...] -r [refgene] + [-f [attrID]] [--rRNA [rRNA]] -o [dir] [--stranded [stranded]] + [-q [mapq]] [-l labels [labels ...]] [-t NUMTHREADS] + +optional arguments: + +:: + + -h, --help show this help message and exit. + -i, --inputFile alignment files. Could be multiple SAM/BAM files separated by space. Required. + -r, --refgene gene annotation file in GTF format. Required + -f the read summation at which feature level in the GTF file. DEFAULT: gene_id. + --rRNA rRNA coordinates in BED format. + -o, --outputDir output directory. Required. + --stranded strandness of the library? + yes : sense stranded + reverse : reverse stranded + no : not stranded + DEFAULT: yes. + -q, --mapq Minimum mapping quality (phred scaled) for an alignment to be called uniquely mapped. DEFAULT:30 + -l, --label Labels of input files. DEFAULT:smp1 smp2 ... + -t, --threads Number of threads to use. DEFAULT:1 + +Example: + +:: + + ezBAMQC -i test-data/exp_data/treat1.bam test-data/exp_data/treat2.bam test-data/exp_data/treat3.bam -r test-data/exp_data/hg9_refGene.gtf -q 30 --rRNA test-data/exp_data/hg19_rRNA.bed -o exp_output2 + + Please find the example output from folder test-data. + +FAQ +=== +Q: Why use ezBAMQC? + +A: ezBAMQC is efficient and easy to use. With one command line, it reports a comprehensive evaluation of the data with a set of plots and tables.The ability to assess multiple samples together with high efficiency make it especially useful in cases where there are a large number of samples from the same condition, genotype, or treatment. ezBAMQC was written in C++ and supports multithreading. A mouse RNA-seq sample with 120M alignments can be done in 8 minutes with 5 threads. + +Q: Why the total number of reads reported by ezBAMQC does not match with samtools flagstat? + +A: The difference is because of non-uniquely mapped reads or multiply aligned reads (multi-reads). Samtools flagstat counts each multiple aligment as a different reads, but ezBAMQC counts reads accoriding to the read ID, i.e., each individual read will be counted once no matter that it is a uniquely mapped read or multi-read. + +Q: What is "Low Quality Reads" ? + +A: Reads marked as qc fail accoriding to SAM format or reads with mapping quality lower than the value set by the option -q will be considered as "Low Quality Reads". + +Q: How the setting of option -q alter the results? + +A: Reads with low quality, i.e., did not pass -q cutoff, are only counted in Total Reads, Mapped Reads, and Mappability by mapping quality plot. The rest of the report does not include low quality reads. + +Q: Do multi-reads (non-uniquely mapped reads) have been considered in Read distribution and gene quantification? + +A: No. Only uniquely mapped reads were counted. + + +Acknowledgements +================ + +#) Samtools contributors +#) Users' valuable feedback + +Copying & Distribution +====================== + +ezBAMQC is free software: you can redistribute it and/or modify +it under the terms of the GNU General Public License as published by +the Free Software Foundation, either version 3 of the License, or +(at your option) any later version. + +This program is distributed in the hope that it will be useful, +but *WITHOUT ANY WARRANTY*; without even the implied warranty of +*MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE*. See the +GNU General Public License for more details. + +You should have received a copy of the GNU General Public License +along with ezBAMQC. If not, see `this website <http://www.gnu.org/licenses/>`_