I'm trying to analyse my rna-seq data with irap.
However, I got some errors during the analysis.
Please, give me some solutions and help me.
Very many thanks.
----------------------------
[irap@localhost ~]$ irap conf=human_ex.conf de_method=deseq
*****************************************************
* IRAP 0.4.0
* Developed by Nuno Fonseca (authorname (at) acm.org)
* This pipeline is distributed under the terms of the GNU General Public License 3
*
* Initializing...
* Trying to load configuration file human_ex.conf...
* Configuration loaded.
*
* Required Parameters:
* name=human_ex
* data_dir=/run/media/irap/KSH/Human
* species=homo_sapiens
* reference=Homo_sapiens.GRCh37.74.dna.toplevel.fa.gz
* gtf_file = Homo_sapiens.GRCh37.74.gtf.gz
* gff3_file = Homo_sapiens.GRCh37.74.gff3
* Transcripts = /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.cdna.all.fa
* pe=myLib1 myLib2 myLib3 myLib4 myLib5
* debug * fastq_files=/home/irap/730-mono-TSP1_1.fastq /home/irap/730-mono-TSP1_2.fastq /home/irap/730-M1-TSP1_1.fastq /home/irap/730-M1-TSP1_2.fastq /home/irap/730-M2a-TSP1_1.fastq /home/irap/730-M2a-TSP1_2.fastq /home/irap/730-M2b-TSP1_1.fastq /home/irap/730-M2b-TSP1_2.fastq /home/irap/730-M2c-TSP1_1.fastq /home/irap/730-M2c-TSP1_2.fastq
* myLib1_sd=60
* myLib2_sd=60
* myLib3_sd=60
* myLib4_sd=60
* myLib5_sd=60
* myLib1_ins=350
* myLib2_ins=350
* myLib3_ins=350
* myLib4_ins=350
* myLib5_ins=350
* myLib1_rs=101
* myLib2_rs=101
* myLib3_rs=101
* myLib4_rs=101
* myLib5_rs=101
* myLib1_qual=33
* myLib2_qual=33
* myLib3_qual=33
* myLib4_qual=33
* myLib5_qual=33
* contrasts=GAvsGB GBvsGA
* GAvsGB=GA GB
* GA=myLib1
* GB=myLib2 myLib3 myLib4 myLib5
* GBvsGA=GB GA
* GB=myLib2 myLib3 myLib4 myLib5
* GA=myLib1
* groups= parameter not defined, using the groups in the contrasts (if defined).
* technical.replicates=NONE
*
* Optional Parameters:
* max_threads=16
* tmp_dir=/run/media/irap/KSH/Human/tmp (temporary directory)
* mapper=tophat1
* quant_method=htseq2
* exon_quant=n
* transcript_quant=y
* quant_norm_method=deseq
* mapper_splicing=yes
* de_method=deseq
* gse_tool=piano
* gse_method=fisher
* gse_pvalue=0.05
* gse_minsize=3 (minimum number of genes)
* gse_minedge=3 (used in the GSE related plots)
* gse_top=Inf (used in the GSE related plots)
*========================================================
DEBUG
reference_prefix=/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
gtf_file_abspath=/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf
index_files= /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.1.ebwt
file_indexed=/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
17:14:02 14/01/2015 * [DONE] Initialization
17:14:02 14/01/2015 * [DONE] Directory structure created
gunzip -c /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gz > /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.tmp && mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.tmp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf
gtf2geneclass.sh /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf > /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gene_class.txt.tmp && mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gene_class.txt.tmp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gene_class.txt
cp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gene_class.txt human_ex/data/gene_class.txt.tmp && mv human_ex/data/gene_class.txt.tmp human_ex/data/gene_class.txt
gunzip -c /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.gz > /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.tmp && mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.tmp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
19:41:31 14/01/2015 * INDEXING
irap_map.sh bowtie1 bowtie-build --offrate 3 /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
Wrapper: bowtie-build --offrate 3 /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
Settings:
Output files: "/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.*.ebwt"
Line rate: 6 (line is 64 bytes)
Lines per side: 1 (side is 64 bytes)
Offset rate: 3 (one in 8)
FTable chars: 10
Strings: unpacked
Max bucket size: default
Max bucket size, sqrt multiplier: default
Max bucket size, len divisor: 4
Difference-cover sample period: 1024
Endianness: little
Actual local endianness: little
Sanity checking: disabled
Assertions: disabled
Random seed: 0
Sizeofs: void*:8, int:4, long:8, size_t:8
Input files DNA, FASTA:
/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
Reading reference sizes
Time reading reference sizes: 02:57:34
Calculating joined length
Writing header
Reserving space for joined string
Joining reference sequences
Time to join reference sequences: 02:25:43
bmax according to bmaxDivN setting: 747254778
Using parameters --bmax 560441084 --dcv 1024
Doing ahead-of-time memory usage test
Passed! Constructing with these parameters: --bmax 560441084 --dcv 1024
Constructing suffix-array element generator
Building DifferenceCoverSample
Building sPrime
Building sPrimeOrder
V-Sorting samples
V-Sorting samples time: 00:03:21
Allocating rank array
Ranking v-sort output
Ranking v-sort output time: 00:00:39
Invoking Larsson-Sadakane on ranks
Invoking Larsson-Sadakane on ranks time: 00:01:34
Sanity-checking and returning
Building samples
Reserving space for 12 sample suffixes
Generating random suffixes
QSorting 12 sample offsets, eliminating duplicates
QSorting sample offsets, eliminating duplicates time: 00:00:00
Multikey QSorting 12 samples
(Using difference cover)
Multikey QSorting samples time: 00:00:00
Calculating bucket sizes
Binary sorting into buckets
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Binary sorting into buckets time: 00:02:20
Splitting and merging
Splitting and merging time: 00:00:00
Split 1, merged 6; iterating...
Binary sorting into buckets
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Binary sorting into buckets time: 00:01:59
Splitting and merging
Splitting and merging time: 00:00:00
Avg bucket size: 3.73627e+08 (target: 560441083)
Converting suffix-array elements to index image
Allocating ftab, absorbFtab
Entering Ebwt loop
Getting block 1 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:39
Sorting block of length 532692084
(Using difference cover)
Sorting block time: 00:13:20
Returning block of 532692085
Getting block 2 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:45
Sorting block of length 305168893
(Using difference cover)
Sorting block time: 00:07:44
Returning block of 305168894
Getting block 3 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:52
Sorting block of length 497342339
(Using difference cover)
Sorting block time: 00:13:34
Returning block of 497342340
Getting block 4 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:54
Sorting block of length 326395284
(Using difference cover)
Sorting block time: 00:08:55
Returning block of 326395285
Getting block 5 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:55
Sorting block of length 336824268
(Using difference cover)
Sorting block time: 00:09:55
Returning block of 336824269
Getting block 6 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:55
Sorting block of length 382684617
(Using difference cover)
Sorting block time: 00:09:40
Returning block of 382684618
Getting block 7 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:54
Sorting block of length 496777301
(Using difference cover)
Sorting block time: 00:12:57
Returning block of 496777302
Getting block 8 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:32
Sorting block of length 111134319
(Using difference cover)
Sorting block time: 00:02:57
Returning block of 111134320
Exited Ebwt loop
fchr[A]: 0
fchr[C]: 881098029
fchr[G]: 1493792045
fchr[T]: 2106926216
fchr[$]: 2989019112
Exiting Ebwt::buildToDisk()
Returning from initFromVector
Wrote 858235346 bytes to primary EBWT file: /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.1.ebwt
Wrote 1494509564 bytes to secondary EBWT file: /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.2.ebwt
Re-opening _in1 and _in2 as input streams
Returning from Ebwt constructor
Headers:
len: 2989019112
bwtLen: 2989019113
sz: 747254778
bwtSz: 747254779
lineRate: 6
linesPerSide: 1
offRate: 3
offMask: 0xfffffff8
isaRate: -1
isaMask: 0xffffffff
ftabChars: 10
eftabLen: 20
eftabSz: 80
ftabLen: 1048577
ftabSz: 4194308
offsLen: 373627390
offsSz: 1494509560
isaLen: 0
isaSz: 0
lineSz: 64
sideSz: 64
sideBwtSz: 56
sideBwtLen: 224
numSidePairs: 6671918
numSides: 13343836
numLines: 13343836
ebwtTotLen: 854005504
ebwtTotSz: 854005504
reverse: 0
Total time for call to driver() for forward index: 07:16:50
Reading reference sizes
Time reading reference sizes: 02:03:56
Calculating joined length
Writing header
Reserving space for joined string
Joining reference sequences
Time to join reference sequences: 02:02:31
bmax according to bmaxDivN setting: 747254778
Using parameters --bmax 560441084 --dcv 1024
Doing ahead-of-time memory usage test
Passed! Constructing with these parameters: --bmax 560441084 --dcv 1024
Constructing suffix-array element generator
Building DifferenceCoverSample
Building sPrime
Building sPrimeOrder
V-Sorting samples
V-Sorting samples time: 00:03:17
Allocating rank array
Ranking v-sort output
Ranking v-sort output time: 00:00:39
Invoking Larsson-Sadakane on ranks
Invoking Larsson-Sadakane on ranks time: 00:01:38
Sanity-checking and returning
Building samples
Reserving space for 12 sample suffixes
Generating random suffixes
QSorting 12 sample offsets, eliminating duplicates
QSorting sample offsets, eliminating duplicates time: 00:00:00
Multikey QSorting 12 samples
(Using difference cover)
Multikey QSorting samples time: 00:00:00
Calculating bucket sizes
Binary sorting into buckets
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Binary sorting into buckets time: 00:02:21
Splitting and merging
Splitting and merging time: 00:00:00
Avg bucket size: 3.73627e+08 (target: 560441083)
Converting suffix-array elements to index image
Allocating ftab, absorbFtab
Entering Ebwt loop
Getting block 1 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:32
Sorting block of length 269521119
(Using difference cover)
Sorting block time: 00:06:28
Returning block of 269521120
Getting block 2 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:43
Sorting block of length 539929538
(Using difference cover)
Sorting block time: 00:13:42
Returning block of 539929539
Getting block 3 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:51
Sorting block of length 470436118
(Using difference cover)
Sorting block time: 00:12:15
Returning block of 470436119
Getting block 4 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:53
Sorting block of length 344556343
(Using difference cover)
Sorting block time: 00:08:54
Returning block of 344556344
Getting block 5 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:49
Sorting block of length 287564152
(Using difference cover)
Sorting block time: 00:07:58
Returning block of 287564153
Getting block 6 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:54
Sorting block of length 317250824
(Using difference cover)
Sorting block time: 00:07:52
Returning block of 317250825
Getting block 7 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:53
Sorting block of length 457404276
(Using difference cover)
Sorting block time: 00:11:34
Returning block of 457404277
Getting block 8 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:36
Sorting block of length 302356735
(Using difference cover)
Sorting block time: 00:07:35
Returning block of 302356736
Exited Ebwt loop
fchr[A]: 0
fchr[C]: 881098029
fchr[G]: 1493792045
fchr[T]: 2106926216
fchr[$]: 2989019112
Exiting Ebwt::buildToDisk()
Returning from initFromVector
Wrote 858235346 bytes to primary EBWT file: /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.rev.1.ebwt
Wrote 1494509564 bytes to secondary EBWT file: /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.rev.2.ebwt
Re-opening _in1 and _in2 as input streams
Returning from Ebwt constructor
Headers:
len: 2989019112
bwtLen: 2989019113
sz: 747254778
bwtSz: 747254779
lineRate: 6
linesPerSide: 1
offRate: 3
offMask: 0xfffffff8
isaRate: -1
isaMask: 0xffffffff
ftabChars: 10
eftabLen: 20
eftabSz: 80
ftabLen: 1048577
ftabSz: 4194308
offsLen: 373627390
offsSz: 1494509560
isaLen: 0
isaSz: 0
lineSz: 64
sideSz: 64
sideBwtSz: 56
sideBwtLen: 224
numSidePairs: 6671918
numSides: 13343836
numLines: 13343836
ebwtTotLen: 854005504
ebwtTotSz: 854005504
reverse: 0
Total time for backward call to driver() for mirror index: 05:54:11
irap_check_gtf /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf && touch /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.checked
Features found: CDS exon start_codon stop_codon
File '/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf' looks ok.
gtf_add_exon_id.pl /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf > /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.exon_id.gtf.tmp && mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.exon_id.gtf.tmp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.exon_id.gtf
Loading /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf
tophat2_gtf_juncs /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf > human_ex/data/Homo_sapiens.GRCh37.74.gtf.juncs.tmp
mv human_ex/data/Homo_sapiens.GRCh37.74.gtf.juncs.tmp human_ex/data/Homo_sapiens.GRCh37.74.gtf.juncs
gtf_juncs v2.0.9 (3067M)
---------------------------
Extracted 375869 junctions from /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf
irap_gtf2mapping --gtf /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf --out /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.tmp --cores 16 && \
mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.tmp.Rdata /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.Rdata && \
mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.tmp_exons.tsv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping_exons.tsv && \
mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.tmp_trans.tsv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping_trans.tsv
Error in `$<-.data.frame`(`*tmp*`, "source", value = c("processed_transcript;transcribed_unprocessed_pseudogene", :
replacement has 234790 rows, data has 1306656
Calls: $<- -> $<-.data.frame
Execution halted
make: *** [/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.Rdata] Error 1
However, I got some errors during the analysis.
Please, give me some solutions and help me.
Very many thanks.
----------------------------
[irap@localhost ~]$ irap conf=human_ex.conf de_method=deseq
*****************************************************
* IRAP 0.4.0
* Developed by Nuno Fonseca (authorname (at) acm.org)
* This pipeline is distributed under the terms of the GNU General Public License 3
*
* Initializing...
* Trying to load configuration file human_ex.conf...
* Configuration loaded.
*
* Required Parameters:
* name=human_ex
* data_dir=/run/media/irap/KSH/Human
* species=homo_sapiens
* reference=Homo_sapiens.GRCh37.74.dna.toplevel.fa.gz
* gtf_file = Homo_sapiens.GRCh37.74.gtf.gz
* gff3_file = Homo_sapiens.GRCh37.74.gff3
* Transcripts = /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.cdna.all.fa
* pe=myLib1 myLib2 myLib3 myLib4 myLib5
* debug * fastq_files=/home/irap/730-mono-TSP1_1.fastq /home/irap/730-mono-TSP1_2.fastq /home/irap/730-M1-TSP1_1.fastq /home/irap/730-M1-TSP1_2.fastq /home/irap/730-M2a-TSP1_1.fastq /home/irap/730-M2a-TSP1_2.fastq /home/irap/730-M2b-TSP1_1.fastq /home/irap/730-M2b-TSP1_2.fastq /home/irap/730-M2c-TSP1_1.fastq /home/irap/730-M2c-TSP1_2.fastq
* myLib1_sd=60
* myLib2_sd=60
* myLib3_sd=60
* myLib4_sd=60
* myLib5_sd=60
* myLib1_ins=350
* myLib2_ins=350
* myLib3_ins=350
* myLib4_ins=350
* myLib5_ins=350
* myLib1_rs=101
* myLib2_rs=101
* myLib3_rs=101
* myLib4_rs=101
* myLib5_rs=101
* myLib1_qual=33
* myLib2_qual=33
* myLib3_qual=33
* myLib4_qual=33
* myLib5_qual=33
* contrasts=GAvsGB GBvsGA
* GAvsGB=GA GB
* GA=myLib1
* GB=myLib2 myLib3 myLib4 myLib5
* GBvsGA=GB GA
* GB=myLib2 myLib3 myLib4 myLib5
* GA=myLib1
* groups= parameter not defined, using the groups in the contrasts (if defined).
* technical.replicates=NONE
*
* Optional Parameters:
* max_threads=16
* tmp_dir=/run/media/irap/KSH/Human/tmp (temporary directory)
* mapper=tophat1
* quant_method=htseq2
* exon_quant=n
* transcript_quant=y
* quant_norm_method=deseq
* mapper_splicing=yes
* de_method=deseq
* gse_tool=piano
* gse_method=fisher
* gse_pvalue=0.05
* gse_minsize=3 (minimum number of genes)
* gse_minedge=3 (used in the GSE related plots)
* gse_top=Inf (used in the GSE related plots)
*========================================================
DEBUG
reference_prefix=/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
gtf_file_abspath=/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf
index_files= /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.1.ebwt
file_indexed=/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
17:14:02 14/01/2015 * [DONE] Initialization
17:14:02 14/01/2015 * [DONE] Directory structure created
gunzip -c /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gz > /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.tmp && mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.tmp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf
gtf2geneclass.sh /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf > /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gene_class.txt.tmp && mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gene_class.txt.tmp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gene_class.txt
cp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.gene_class.txt human_ex/data/gene_class.txt.tmp && mv human_ex/data/gene_class.txt.tmp human_ex/data/gene_class.txt
gunzip -c /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.gz > /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.tmp && mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.tmp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
19:41:31 14/01/2015 * INDEXING
irap_map.sh bowtie1 bowtie-build --offrate 3 /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
Wrapper: bowtie-build --offrate 3 /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
Settings:
Output files: "/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.*.ebwt"
Line rate: 6 (line is 64 bytes)
Lines per side: 1 (side is 64 bytes)
Offset rate: 3 (one in 8)
FTable chars: 10
Strings: unpacked
Max bucket size: default
Max bucket size, sqrt multiplier: default
Max bucket size, len divisor: 4
Difference-cover sample period: 1024
Endianness: little
Actual local endianness: little
Sanity checking: disabled
Assertions: disabled
Random seed: 0
Sizeofs: void*:8, int:4, long:8, size_t:8
Input files DNA, FASTA:
/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa
Reading reference sizes
Time reading reference sizes: 02:57:34
Calculating joined length
Writing header
Reserving space for joined string
Joining reference sequences
Time to join reference sequences: 02:25:43
bmax according to bmaxDivN setting: 747254778
Using parameters --bmax 560441084 --dcv 1024
Doing ahead-of-time memory usage test
Passed! Constructing with these parameters: --bmax 560441084 --dcv 1024
Constructing suffix-array element generator
Building DifferenceCoverSample
Building sPrime
Building sPrimeOrder
V-Sorting samples
V-Sorting samples time: 00:03:21
Allocating rank array
Ranking v-sort output
Ranking v-sort output time: 00:00:39
Invoking Larsson-Sadakane on ranks
Invoking Larsson-Sadakane on ranks time: 00:01:34
Sanity-checking and returning
Building samples
Reserving space for 12 sample suffixes
Generating random suffixes
QSorting 12 sample offsets, eliminating duplicates
QSorting sample offsets, eliminating duplicates time: 00:00:00
Multikey QSorting 12 samples
(Using difference cover)
Multikey QSorting samples time: 00:00:00
Calculating bucket sizes
Binary sorting into buckets
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Binary sorting into buckets time: 00:02:20
Splitting and merging
Splitting and merging time: 00:00:00
Split 1, merged 6; iterating...
Binary sorting into buckets
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Binary sorting into buckets time: 00:01:59
Splitting and merging
Splitting and merging time: 00:00:00
Avg bucket size: 3.73627e+08 (target: 560441083)
Converting suffix-array elements to index image
Allocating ftab, absorbFtab
Entering Ebwt loop
Getting block 1 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:39
Sorting block of length 532692084
(Using difference cover)
Sorting block time: 00:13:20
Returning block of 532692085
Getting block 2 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:45
Sorting block of length 305168893
(Using difference cover)
Sorting block time: 00:07:44
Returning block of 305168894
Getting block 3 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:52
Sorting block of length 497342339
(Using difference cover)
Sorting block time: 00:13:34
Returning block of 497342340
Getting block 4 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:54
Sorting block of length 326395284
(Using difference cover)
Sorting block time: 00:08:55
Returning block of 326395285
Getting block 5 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:55
Sorting block of length 336824268
(Using difference cover)
Sorting block time: 00:09:55
Returning block of 336824269
Getting block 6 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:55
Sorting block of length 382684617
(Using difference cover)
Sorting block time: 00:09:40
Returning block of 382684618
Getting block 7 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:54
Sorting block of length 496777301
(Using difference cover)
Sorting block time: 00:12:57
Returning block of 496777302
Getting block 8 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:32
Sorting block of length 111134319
(Using difference cover)
Sorting block time: 00:02:57
Returning block of 111134320
Exited Ebwt loop
fchr[A]: 0
fchr[C]: 881098029
fchr[G]: 1493792045
fchr[T]: 2106926216
fchr[$]: 2989019112
Exiting Ebwt::buildToDisk()
Returning from initFromVector
Wrote 858235346 bytes to primary EBWT file: /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.1.ebwt
Wrote 1494509564 bytes to secondary EBWT file: /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.2.ebwt
Re-opening _in1 and _in2 as input streams
Returning from Ebwt constructor
Headers:
len: 2989019112
bwtLen: 2989019113
sz: 747254778
bwtSz: 747254779
lineRate: 6
linesPerSide: 1
offRate: 3
offMask: 0xfffffff8
isaRate: -1
isaMask: 0xffffffff
ftabChars: 10
eftabLen: 20
eftabSz: 80
ftabLen: 1048577
ftabSz: 4194308
offsLen: 373627390
offsSz: 1494509560
isaLen: 0
isaSz: 0
lineSz: 64
sideSz: 64
sideBwtSz: 56
sideBwtLen: 224
numSidePairs: 6671918
numSides: 13343836
numLines: 13343836
ebwtTotLen: 854005504
ebwtTotSz: 854005504
reverse: 0
Total time for call to driver() for forward index: 07:16:50
Reading reference sizes
Time reading reference sizes: 02:03:56
Calculating joined length
Writing header
Reserving space for joined string
Joining reference sequences
Time to join reference sequences: 02:02:31
bmax according to bmaxDivN setting: 747254778
Using parameters --bmax 560441084 --dcv 1024
Doing ahead-of-time memory usage test
Passed! Constructing with these parameters: --bmax 560441084 --dcv 1024
Constructing suffix-array element generator
Building DifferenceCoverSample
Building sPrime
Building sPrimeOrder
V-Sorting samples
V-Sorting samples time: 00:03:17
Allocating rank array
Ranking v-sort output
Ranking v-sort output time: 00:00:39
Invoking Larsson-Sadakane on ranks
Invoking Larsson-Sadakane on ranks time: 00:01:38
Sanity-checking and returning
Building samples
Reserving space for 12 sample suffixes
Generating random suffixes
QSorting 12 sample offsets, eliminating duplicates
QSorting sample offsets, eliminating duplicates time: 00:00:00
Multikey QSorting 12 samples
(Using difference cover)
Multikey QSorting samples time: 00:00:00
Calculating bucket sizes
Binary sorting into buckets
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Binary sorting into buckets time: 00:02:21
Splitting and merging
Splitting and merging time: 00:00:00
Avg bucket size: 3.73627e+08 (target: 560441083)
Converting suffix-array elements to index image
Allocating ftab, absorbFtab
Entering Ebwt loop
Getting block 1 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:32
Sorting block of length 269521119
(Using difference cover)
Sorting block time: 00:06:28
Returning block of 269521120
Getting block 2 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:43
Sorting block of length 539929538
(Using difference cover)
Sorting block time: 00:13:42
Returning block of 539929539
Getting block 3 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:51
Sorting block of length 470436118
(Using difference cover)
Sorting block time: 00:12:15
Returning block of 470436119
Getting block 4 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:53
Sorting block of length 344556343
(Using difference cover)
Sorting block time: 00:08:54
Returning block of 344556344
Getting block 5 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:49
Sorting block of length 287564152
(Using difference cover)
Sorting block time: 00:07:58
Returning block of 287564153
Getting block 6 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:54
Sorting block of length 317250824
(Using difference cover)
Sorting block time: 00:07:52
Returning block of 317250825
Getting block 7 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:53
Sorting block of length 457404276
(Using difference cover)
Sorting block time: 00:11:34
Returning block of 457404277
Getting block 8 of 8
Reserving size (560441084) for bucket
Calculating Z arrays
Calculating Z arrays time: 00:00:00
Entering block accumulator loop:
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Block accumulator loop time: 00:00:36
Sorting block of length 302356735
(Using difference cover)
Sorting block time: 00:07:35
Returning block of 302356736
Exited Ebwt loop
fchr[A]: 0
fchr[C]: 881098029
fchr[G]: 1493792045
fchr[T]: 2106926216
fchr[$]: 2989019112
Exiting Ebwt::buildToDisk()
Returning from initFromVector
Wrote 858235346 bytes to primary EBWT file: /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.rev.1.ebwt
Wrote 1494509564 bytes to secondary EBWT file: /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.dna.toplevel.fa.rev.2.ebwt
Re-opening _in1 and _in2 as input streams
Returning from Ebwt constructor
Headers:
len: 2989019112
bwtLen: 2989019113
sz: 747254778
bwtSz: 747254779
lineRate: 6
linesPerSide: 1
offRate: 3
offMask: 0xfffffff8
isaRate: -1
isaMask: 0xffffffff
ftabChars: 10
eftabLen: 20
eftabSz: 80
ftabLen: 1048577
ftabSz: 4194308
offsLen: 373627390
offsSz: 1494509560
isaLen: 0
isaSz: 0
lineSz: 64
sideSz: 64
sideBwtSz: 56
sideBwtLen: 224
numSidePairs: 6671918
numSides: 13343836
numLines: 13343836
ebwtTotLen: 854005504
ebwtTotSz: 854005504
reverse: 0
Total time for backward call to driver() for mirror index: 05:54:11
irap_check_gtf /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf && touch /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.checked
Features found: CDS exon start_codon stop_codon
File '/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf' looks ok.
gtf_add_exon_id.pl /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf > /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.exon_id.gtf.tmp && mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.exon_id.gtf.tmp /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf.exon_id.gtf
Loading /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf
tophat2_gtf_juncs /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf > human_ex/data/Homo_sapiens.GRCh37.74.gtf.juncs.tmp
mv human_ex/data/Homo_sapiens.GRCh37.74.gtf.juncs.tmp human_ex/data/Homo_sapiens.GRCh37.74.gtf.juncs
gtf_juncs v2.0.9 (3067M)
---------------------------
Extracted 375869 junctions from /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf
irap_gtf2mapping --gtf /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.gtf --out /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.tmp --cores 16 && \
mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.tmp.Rdata /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.Rdata && \
mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.tmp_exons.tsv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping_exons.tsv && \
mv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.tmp_trans.tsv /run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping_trans.tsv
Error in `$<-.data.frame`(`*tmp*`, "source", value = c("processed_transcript;transcribed_unprocessed_pseudogene", :
replacement has 234790 rows, data has 1306656
Calls: $<- -> $<-.data.frame
Execution halted
make: *** [/run/media/irap/KSH/Human/reference/homo_sapiens/Homo_sapiens.GRCh37.74.mapping.Rdata] Error 1
Comment