BSMAP-methatio-methykit workflow

In [10]:
#Setting Variables
#file ID
fid="CgLarv_T1D5_sept"
#where is bsmap
bsmap="/Users/Shared/Apps/bsmap-2.73/"
#fastq files location R1 location
R1="/Volumes/web/trilobite/Crassostrea_gigas_HTSdata/batterbox/BiGo_Larvae/filtered_BS_CgLarv_T1D5_ACAGTG_L007_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"
#location of sqlshare python client tools
spt="/Users/Mackenzie/sqlshare-pythonclient/tools/"
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_T1D5_sept

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

BSMAP v2.73
Start at:  Wed Feb 12 17:22:20 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. 8 secs passed
total_kmers: 43046721
Create seed table. 25 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/BiGo_Larvae/filtered_BS_CgLarv_T1D5_ACAGTG_L007_R1.fastq.gz 	(format: gzipped FASTQ)
Output file: bsmap_out.sam	 (format: SAM)
Thread #0: 	50000 reads finished. 30 secs passed
Thread #0: 	100000 reads finished. 35 secs passed
Thread #0: 	150000 reads finished. 39 secs passed
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Thread #0: 	2700000 reads finished. 289 secs passed
Thread #0: 	2709822 reads finished. 290 secs passed
Total number of aligned reads: 1905030 (70%)
Done.
Finished at Wed Feb 12 17:27:10 2014
Total time consumed:  290 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 17:27:11 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 17:27:45 2014: reading bsmap_out.sam ...
[samopen] SAM header is present: 7658 sequences.
@ Wed Feb 12 17:28:49 2014: combining CpG methylation from both strands ...
@ Wed Feb 12 17:29:20 2014: writing methratio_out.txt ...
@ Wed Feb 12 17:33:44 2014: done.
total 1454458 valid mappings, 13898482 covered cytosines, average coverage: 1.16 fold.

In [11]:
 !python {spt}singleupload.py -u che625@washington.edu -p 5234162537ce6a35236569c28ab62f65 -d _methratio{fid} methratio_out.txt 
processing chunk line 0 to 1833148 (1.93166208267 s elapsed)
pushing methratio_out.txt...
parsing AA9140E1...
processing chunk line 1833148 to 3626707 (719.435286999 s elapsed)
pushing methratio_out.txt...
parsing 918A1FB8...
processing chunk line 3626707 to 5420385 (1481.80443597 s elapsed)
pushing methratio_out.txt...
parsing 4CE37E82...
processing chunk line 5420385 to 7232566 (2262.96288896 s elapsed)
pushing methratio_out.txt...
parsing F572D180...
processing chunk line 7232566 to 9022469 (2956.26540112 s elapsed)
pushing methratio_out.txt...
parsing A9D6C699...
processing chunk line 9022469 to 10802862 (3444.55271196 s elapsed)
pushing methratio_out.txt...
parsing 83DD5AB8...
processing chunk line 10802862 to 12623989 (3935.587538 s elapsed)
pushing methratio_out.txt...
parsing BF817988...
processing chunk line 12623989 to 13898483 (4312.01511002 s elapsed)
pushing methratio_out.txt...
parsing 5F38B432...
finished _methratioCgLarv_T1D5_sept

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