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Alignment of bisulfite sequence reads and tabulation of read-level methylation measurements
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fumi-github/bsmooth-align
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BSmooth is by Kasper D. Hansen, Ben Langmead, Rafael A. Irizarry Statistics pipeline is by Kasper D. Hansen Alignment pipeline is by Ben Langmead The BSmooth alignment pipeline is licensed under the GPLv3 license. See `LICENSE' file for details. Installation ============ BSmooth ------- To install the BSmooth alignment pipeline, permanently set the BSMOOTH_HOME environment variable to the main BSmooth directory. I.e., the directory with 'bin', 'example' and 'lib' subdirectories inside. Building merman --------------- The merman sources are included with BSmooth. To use merman with BSmooth, you must first build the merman binary. This requires standard GNU build tools such as GNU make and g++. To build merman, cd to the $BSMOOTH_HOME/merman directory and run 'make'. Finally, add the $BSMOOTH_HOME/merman directory to your PATH. Installing Bowtie 2 ------------------- Follow the standard instructions for installing those tools; see their respective manuals for details. Once the tool is installed, make sure that the directory containing the key binaries ('bowtie2', 'bowtie2-build') is included in your PATH. Pipeline flow ============= The following 4 steps (or possibly 3 steps, depending on aligner) are needed to get from raw input reads to CpG-level measurement summaries. These summaries constitute the input to the R pipeline for smoothing and calculating DMRs. Step 1: Build bisulfite genome index (not necessary if using Merman) bswc_bowtie2_index.pl Step 2: Align bs_merman_align.pl bswc_bowtie2_align.pl Step 3: Sort evidence directory bsev_sort.pl Step 4: Tabulate sorted evidence directory bsev_tabulate.pl For an example, see example/Makefile, which uses example/sim.pl to create a small, simulated problem, then goes through the steps listed above.
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