BSMAP-methatio-methykit workflow

In [13]:
#Setting Variables
#file ID
fid="CgLarv_T1D3_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_T1D3_TGACCA_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 [8]:
cd /Volumes/web/Mollusk/bs_larvae_exp/
/Volumes/web/Mollusk/bs_larvae_exp

In [9]:
mkdir {fid}
In [10]:
cd {fid}
/Volumes/web/Mollusk/bs_larvae_exp/CgLarv_T1D3_sept

In [11]:
!{bsmap}bsmap -a {R1} -d {genome} -o bsmap_out.sam -p 1

BSMAP v2.73
Start at:  Wed Feb 12 17:08:33 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. 17 secs passed
total_kmers: 43046721
Create seed table. 32 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_T1D3_TGACCA_L007_R1.fastq.gz 	(format: gzipped FASTQ)
Output file: bsmap_out.sam	 (format: SAM)
Thread #0: 	50000 reads finished. 37 secs passed
Thread #0: 	100000 reads finished. 42 secs passed
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Thread #0: 	3343987 reads finished. 355 secs passed
Total number of aligned reads: 2346435 (70%)
Done.
Finished at Wed Feb 12 17:14:28 2014
Total time consumed:  355 secs

In [12]:
!python {bsmap}methratio.py -d {genome} -u -z -g -o methratio_out.txt -s {bsmap}samtools bsmap_out.sam
@ Wed Feb 12 17:14:28 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 17:15:02 2014: reading bsmap_out.sam ...
[samopen] SAM header is present: 7658 sequences.
@ Wed Feb 12 17:16:23 2014: combining CpG methylation from both strands ...
@ Wed Feb 12 17:16:53 2014: writing methratio_out.txt ...
@ Wed Feb 12 17:21:32 2014: done.
total 1771949 valid mappings, 16034155 covered cytosines, average coverage: 1.21 fold.

In [14]:
!python {spt}singleupload.py -u che625@washington.edu -p 5234162537ce6a35236569c28ab62f65 -d _methratio{fid} methratio_out.txt 
processing chunk line 0 to 1839222 (1.89109706879 s elapsed)
pushing methratio_out.txt...
parsing 80D3A205...
processing chunk line 1839222 to 3633606 (596.911438942 s elapsed)
pushing methratio_out.txt...
parsing D8807C2A...
processing chunk line 3633606 to 5427075 (1114.29850101 s elapsed)
pushing methratio_out.txt...
parsing D7E5D717...
processing chunk line 5427075 to 7233592 (1627.1321559 s elapsed)
pushing methratio_out.txt...
parsing 08D20CA7...
processing chunk line 7233592 to 9036349 (2367.91254187 s elapsed)
pushing methratio_out.txt...
parsing DBF5878D...
processing chunk line 9036349 to 10820268 (3142.22911191 s elapsed)
pushing methratio_out.txt...
parsing 4F7FA24D...
processing chunk line 10820268 to 12605427 (3875.60892487 s elapsed)
pushing methratio_out.txt...
parsing 1DFB33D3...
processing chunk line 12605427 to 14426750 (4568.89165497 s elapsed)
pushing methratio_out.txt...
parsing 412D4840...
processing chunk line 14426750 to 16034156 (5065.92029905 s elapsed)
pushing methratio_out.txt...
parsing C7170052...
finished _methratioCgLarv_T1D3_sept

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