BSMAP-methatio-methykit workflow

In [7]:
#Setting Variables
#file ID
fid="M1_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_CgM1_ACTTGA_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 [11]:
cd /Volumes/web/Mollusk/bs_larvae_exp/
/Volumes/web/Mollusk/bs_larvae_exp

In [12]:
mkdir {fid}
In [13]:
cd {fid}
/Volumes/web/Mollusk/bs_larvae_exp/M1_sept

In [14]:
!{bsmap}bsmap -a {R1} -d {genome} -o bsmap_out.sam -p 1

BSMAP v2.73
Start at:  Wed Feb 12 14:56:40 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. 24 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_CgM1_ACTTGA_L007_R1.fastq.gz 	(format: gzipped FASTQ)
Output file: bsmap_out.sam	 (format: SAM)
Thread #0: 	50000 reads finished. 29 secs passed
Thread #0: 	100000 reads finished. 34 secs passed
Thread #0: 	150000 reads finished. 39 secs passed
Thread #0: 	200000 reads finished. 43 secs passed
Thread #0: 	250000 reads finished. 48 secs passed
Thread #0: 	300000 reads finished. 53 secs passed
Thread #0: 	350000 reads finished. 58 secs passed
Thread #0: 	400000 reads finished. 63 secs passed
Thread #0: 	450000 reads finished. 67 secs passed
Thread #0: 	500000 reads finished. 72 secs passed
Thread #0: 	550000 reads finished. 76 secs passed
Thread #0: 	600000 reads finished. 81 secs passed
Thread #0: 	650000 reads finished. 86 secs passed
Thread #0: 	700000 reads finished. 90 secs passed
Thread #0: 	750000 reads finished. 95 secs passed
Thread #0: 	800000 reads finished. 100 secs passed
Thread #0: 	850000 reads finished. 104 secs passed
Thread #0: 	900000 reads finished. 109 secs passed
Thread #0: 	950000 reads finished. 114 secs passed
Thread #0: 	1000000 reads finished. 118 secs passed
Thread #0: 	1050000 reads finished. 123 secs passed
Thread #0: 	1100000 reads finished. 128 secs passed
Thread #0: 	1150000 reads finished. 132 secs passed
Thread #0: 	1200000 reads finished. 137 secs passed
Thread #0: 	1250000 reads finished. 141 secs passed
Thread #0: 	1300000 reads finished. 146 secs passed
Thread #0: 	1350000 reads finished. 151 secs passed
Thread #0: 	1400000 reads finished. 155 secs passed
Thread #0: 	1450000 reads finished. 160 secs passed
Thread #0: 	1500000 reads finished. 165 secs passed
Thread #0: 	1550000 reads finished. 169 secs passed
Thread #0: 	1600000 reads finished. 174 secs passed
Thread #0: 	1650000 reads finished. 179 secs passed
Thread #0: 	1700000 reads finished. 183 secs passed
Thread #0: 	1750000 reads finished. 188 secs passed
Thread #0: 	1800000 reads finished. 193 secs passed
Thread #0: 	1850000 reads finished. 198 secs passed
Thread #0: 	1900000 reads finished. 202 secs passed
Thread #0: 	1950000 reads finished. 207 secs passed
Thread #0: 	2000000 reads finished. 212 secs passed
Thread #0: 	2050000 reads finished. 217 secs passed
Thread #0: 	2100000 reads finished. 222 secs passed
Thread #0: 	2150000 reads finished. 227 secs passed
Thread #0: 	2200000 reads finished. 231 secs passed
Thread #0: 	2250000 reads finished. 236 secs passed
Thread #0: 	2300000 reads finished. 241 secs passed
Thread #0: 	2350000 reads finished. 246 secs passed
Thread #0: 	2400000 reads finished. 251 secs passed
Thread #0: 	2450000 reads finished. 264 secs passed
Thread #0: 	2500000 reads finished. 268 secs passed
Thread #0: 	2550000 reads finished. 274 secs passed
Thread #0: 	2600000 reads finished. 279 secs passed
Thread #0: 	2650000 reads finished. 289 secs passed
Thread #0: 	2700000 reads finished. 295 secs passed
Thread #0: 	2750000 reads finished. 299 secs passed
Thread #0: 	2800000 reads finished. 304 secs passed
Thread #0: 	2850000 reads finished. 309 secs passed
Thread #0: 	2900000 reads finished. 314 secs passed
Thread #0: 	2950000 reads finished. 322 secs passed
Thread #0: 	3000000 reads finished. 333 secs passed
Thread #0: 	3050000 reads finished. 338 secs passed
Thread #0: 	3100000 reads finished. 343 secs passed
Thread #0: 	3150000 reads finished. 351 secs passed
Thread #0: 	3200000 reads finished. 356 secs passed
Thread #0: 	3250000 reads finished. 363 secs passed
Thread #0: 	3300000 reads finished. 368 secs passed
Thread #0: 	3350000 reads finished. 380 secs passed
Thread #0: 	3400000 reads finished. 385 secs passed
Thread #0: 	3450000 reads finished. 390 secs passed
Thread #0: 	3500000 reads finished. 398 secs passed
Thread #0: 	3550000 reads finished. 403 secs passed
Thread #0: 	3600000 reads finished. 408 secs passed
Thread #0: 	3650000 reads finished. 412 secs passed
Thread #0: 	3700000 reads finished. 418 secs passed
Thread #0: 	3750000 reads finished. 424 secs passed
Thread #0: 	3800000 reads finished. 432 secs passed
Thread #0: 	3850000 reads finished. 437 secs passed
Thread #0: 	3900000 reads finished. 443 secs passed
Thread #0: 	3950000 reads finished. 448 secs passed
Thread #0: 	4000000 reads finished. 455 secs passed
Thread #0: 	4050000 reads finished. 462 secs passed
Thread #0: 	4100000 reads finished. 467 secs passed
Thread #0: 	4150000 reads finished. 474 secs passed
Thread #0: 	4200000 reads finished. 479 secs passed
Thread #0: 	4250000 reads finished. 484 secs passed
Thread #0: 	4300000 reads finished. 489 secs passed
Thread #0: 	4350000 reads finished. 494 secs passed
Thread #0: 	4400000 reads finished. 499 secs passed
Thread #0: 	4450000 reads finished. 504 secs passed
Thread #0: 	4500000 reads finished. 509 secs passed
Thread #0: 	4550000 reads finished. 514 secs passed
Thread #0: 	4600000 reads finished. 519 secs passed
Thread #0: 	4650000 reads finished. 524 secs passed
Thread #0: 	4700000 reads finished. 529 secs passed
Thread #0: 	4750000 reads finished. 534 secs passed
Thread #0: 	4800000 reads finished. 539 secs passed
Thread #0: 	4850000 reads finished. 544 secs passed
Thread #0: 	4900000 reads finished. 550 secs passed
Thread #0: 	4950000 reads finished. 555 secs passed
Thread #0: 	5000000 reads finished. 559 secs passed
Thread #0: 	5050000 reads finished. 565 secs passed
Thread #0: 	5100000 reads finished. 570 secs passed
Thread #0: 	5150000 reads finished. 575 secs passed
Thread #0: 	5200000 reads finished. 580 secs passed
Thread #0: 	5250000 reads finished. 585 secs passed
Thread #0: 	5300000 reads finished. 589 secs passed
Thread #0: 	5350000 reads finished. 594 secs passed
Thread #0: 	5400000 reads finished. 600 secs passed
Thread #0: 	5450000 reads finished. 605 secs passed
Thread #0: 	5500000 reads finished. 611 secs passed
Thread #0: 	5543163 reads finished. 616 secs passed
Total number of aligned reads: 3687702 (67%)
Done.
Finished at Wed Feb 12 15:06:57 2014
Total time consumed:  617 secs

In [15]:
!python {bsmap}methratio.py -d {genome} -u -z -g -o methratio_out.txt -s {bsmap}samtools bsmap_out.sam
@ Wed Feb 12 15:06:57 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 15:07:33 2014: reading bsmap_out.sam ...
[samopen] SAM header is present: 7658 sequences.
@ Wed Feb 12 15:10:09 2014: combining CpG methylation from both strands ...
@ Wed Feb 12 15:10:43 2014: writing methratio_out.txt ...
@ Wed Feb 12 15:21:48 2014: done.
total 2686629 valid mappings, 21854162 covered cytosines, average coverage: 1.35 fold.

In [8]:
!python {spt}singleupload.py -u che625@washington.edu -p 5234162537ce6a35236569c28ab62f65 -d _methratio{fid} methratio_out.txt 
Traceback (most recent call last):
  File "/Users/Mackenzie/sqlshare-pythonclient/tools/singleupload.py", line 42, in <module>
    main()
  File "/Users/Mackenzie/sqlshare-pythonclient/tools/singleupload.py", line 39, in main
    conn.uploadone(args[0], datasetname)
  File "build/bdist.macosx-10.8-intel/egg/sqlshare/__init__.py", line 166, in uploadone
IOError: [Errno 2] No such file or directory: 'methratio_out.txt'

In []: