BSMAP-methatio-methykit workflow

In [10]:
#Setting Variables
#file ID
fid="M3_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_CgM3/filtered_BS_CgM3_GATCAG_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"
#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/M3_nov

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

BSMAP v2.73
Start at:  Thu Feb 13 09:19:12 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. 10 secs passed
total_kmers: 43046721
Create seed table. 28 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_CgM3/filtered_BS_CgM3_GATCAG_L004_R1.fastq.gz 	(format: gzipped FASTQ)
Output file: bsmap_out.sam	 (format: SAM)
Thread #0: 	50000 reads finished. 33 secs passed
Thread #0: 	100000 reads finished. 38 secs passed
Thread #0: 	150000 reads finished. 43 secs passed
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Thread #0: 	2550000 reads finished. 283 secs passed
Thread #0: 	2551374 reads finished. 283 secs passed
Total number of aligned reads: 1784682 (70%)
Done.
Finished at Thu Feb 13 09:23:55 2014
Total time consumed:  283 secs

In [9]:
!python {bsmap}methratio.py -d {genome} -u -z -g -o methratio_out.txt -s {bsmap}samtools bsmap_out.sam
@ Thu Feb 13 09:23:56 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 ...
@ Thu Feb 13 09:24:31 2014: reading bsmap_out.sam ...
[samopen] SAM header is present: 7658 sequences.
@ Thu Feb 13 09:25:43 2014: combining CpG methylation from both strands ...
@ Thu Feb 13 09:26:15 2014: writing methratio_out.txt ...
@ Thu Feb 13 09:30:25 2014: done.
total 1390421 valid mappings, 12886605 covered cytosines, average coverage: 1.17 fold.

In [11]:
!python {spt}singleupload.py -u che625@washington.edu -p 5234162537ce6a35236569c28ab62f65 -d _methratio{fid} methratio_out.txt 
processing chunk line 0 to 1830536 (1.97255396843 s elapsed)
pushing methratio_out.txt...
parsing 1E6977F9...
processing chunk line 1830536 to 3624509 (520.807501078 s elapsed)
pushing methratio_out.txt...
parsing DD920708...
processing chunk line 3624509 to 5422535 (1095.6222229 s elapsed)
pushing methratio_out.txt...
parsing DD924D74...
processing chunk line 5422535 to 7228960 (1657.58009291 s elapsed)
pushing methratio_out.txt...
parsing 94E2FF30...
processing chunk line 7228960 to 9010254 (2369.8491559 s elapsed)
pushing methratio_out.txt...
parsing 31FB6C8A...
processing chunk line 9010254 to 10811306 (3163.14582992 s elapsed)
pushing methratio_out.txt...
parsing 62C757EB...
processing chunk line 10811306 to 12633900 (3911.77813411 s elapsed)
pushing methratio_out.txt...
parsing A87DE126...
processing chunk line 12633900 to 12886606 (4583.32832599 s elapsed)
pushing methratio_out.txt...
parsing BCF03CF1...
finished _methratioM3_nov

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