BSMAP-methatio-methykit workflow

In [4]:
#Setting Variables
#file ID
fid="CgLarv_T3D5_nov"
#where is bsmap
bsmap="/Users/Shared/Apps/bsmap-2.73/"
#fastq files location R1 location
R1="/Volumes/web/trilobite/Crassostrea_gigas_HTSdata/batterbox/FCC39EM/Sample_BS_CgLarv_T3D5/filtered_BS_CgLarv_T3D5_CAGATC_L004_R1.fastq.gz"
#genome file 
genome="/Volumes/web/whale/ensembl/ftp.ensemblgenomes.org/pub/release-21/metazoa/fasta/crassostrea_gigas/dna/Crassostrea_gigas.GCA_000297895.1.21.dna_sm.genome.fa"
In [5]:
cd /Volumes/web/Mollusk/bs_larvae_exp/
/Volumes/web/Mollusk/bs_larvae_exp

In [6]:
mkdir {fid}
In [7]:
cd {fid}
/Volumes/web/Mollusk/bs_larvae_exp/CgLarv_T3D5_nov

In [8]:
!{bsmap}bsmap -a {R1} -d {genome} -o bsmap_out.sam -p 1

BSMAP v2.73
Start at:  Wed Feb 12 17:34:46 2014

Input reference file: /Volumes/web/whale/ensembl/ftp.ensemblgenomes.org/pub/release-21/metazoa/fasta/crassostrea_gigas/dna/Crassostrea_gigas.GCA_000297895.1.21.dna_sm.genome.fa 	(format: FASTA)
Load in 7658 db seqs, total size 557717710 bp. 9 secs passed
total_kmers: 43046721
Create seed table. 29 secs passed
max number of mismatches: read_length * 8% 	max gap size: 0
kmer cut-off ratio:5e-07
max multi-hits: 100	max Ns: 5	seed size: 16	index interval: 4
quality cutoff: 0	base quality char: '!'
min fragment size:28	max fragemt size:500
start from read #1	end at read #4294967295
additional alignment: T in reads => C in reference
mapping strand: ++,-+
Single-end alignment(1 threads)
Input read file: /Volumes/web/trilobite/Crassostrea_gigas_HTSdata/batterbox/FCC39EM/Sample_BS_CgLarv_T3D5/filtered_BS_CgLarv_T3D5_CAGATC_L004_R1.fastq.gz 	(format: gzipped FASTQ)
Output file: bsmap_out.sam	 (format: SAM)
Thread #0: 	50000 reads finished. 34 secs passed
Thread #0: 	100000 reads finished. 39 secs passed
Thread #0: 	150000 reads finished. 44 secs passed
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Thread #0: 	2800000 reads finished. 330 secs passed
Thread #0: 	2850000 reads finished. 336 secs passed
Thread #0: 	2900000 reads finished. 341 secs passed
Thread #0: 	2929159 reads finished. 344 secs passed
Total number of aligned reads: 2048300 (70%)
Done.
Finished at Wed Feb 12 17:40:30 2014
Total time consumed:  344 secs

In [9]:
!python {bsmap}methratio.py -d {genome} -u -z -g -o methratio_out.txt -s {bsmap}samtools bsmap_out.sam
@ Wed Feb 12 17:40:31 2014: reading reference /Volumes/web/whale/ensembl/ftp.ensemblgenomes.org/pub/release-21/metazoa/fasta/crassostrea_gigas/dna/Crassostrea_gigas.GCA_000297895.1.21.dna_sm.genome.fa ...
@ Wed Feb 12 17:41:36 2014: reading bsmap_out.sam ...
[samopen] SAM header is present: 7658 sequences.
@ Wed Feb 12 18:36:28 2014: combining CpG methylation from both strands ...
@ Wed Feb 12 18:39:41 2014: writing methratio_out.txt ...
@ Wed Feb 12 18:44:19 2014: done.
total 1579195 valid mappings, 14637142 covered cytosines, average coverage: 1.18 fold.

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