Hi,
I am running MISO with paired-end reads but it comes up the error message. Just wondering if anyone had the same problem? Tia!
python test_miso.py
Testing conversion of SAM to BAM...
Executing: python ./Projects/tools/MISO/sam_to_bam.py --convert ./Projects/tools/MISO/test-data/sam-data/c2c12.Atp2b1.sam ./Projects/tools/MISO/test-output/sam-output
Converting SAM to BAM...
- Executing: samtools view -Sbh ./Projects/tools/MISO/test-data/sam-data/c2c12.Atp2b1.sam > ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.bam
[samopen] SAM header is present: 35 sequences.
Sorting BAM file...
- Executing: samtools sort ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.bam ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted
Indexing BAM...
- Executing: samtools index ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam
Conversion took 0.00 minutes.
.Testing single-end SE event interface...
Executing: python ./Projects/tools/MISO/run_events_analysis.py --compute-events-psi se-sample ./Projects/tools/MISO/test-data/se-counts/se_test.counts --output-dir ./Projects/tools/MISO/test-output/SE-output --read-len 35 --overhang-len 4 --event-type SE
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
Loading settings file from: ./Projects/tools/MISO/settings/miso_settings.txt
Computing Psi for events...
Filter events?
- yes
Computing Psi for events of type SE
- samples used: ['se-sample']
Processing sample: label=se-sample, filename=./Projects/tools/MISO/test-data/se-counts/se_test.counts
Filtering events...
Filtering SE events...
Running on a total of 2 events.
Serializing a total of 2 events by Pickle.
Executing: python ./Projects/tools/MISO/run_miso.py --compute-two-iso-psi ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/pickle/se-sample.pickle ./Projects/tools/MISO/test-output/SE-output/SE/se-sample --event-type SE --read-len 35 --overhang-len 4
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
Running MISO on: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/pickle/se-sample.pickle
- Output directory: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/
Called on: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/pickle/se-sample.pickle
Loading from Pickle file took 0.00 seconds.
Parsing of events to genes took 0.00 seconds.
Running sampler on a two-isoform event...
- Gene label: chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:- gene_id: chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:-
isoforms: [Isoform(gene = chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:-, g_start = 0, g_end = 811, len = 812,
parts = ['A', 'B', 'C']), Isoform(gene = chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:-, g_start = 0, g_end = 811, len = 745,
parts = ['A', 'C'])]
- NI, NE, NB: 4, 42, 281
Using default sampler parameters.
Indexing by chromosome...
Output filename ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/chr15/chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:-.miso exists, not running MISO.
Parsing of events to genes took 0.00 seconds.
Running sampler on a two-isoform event...
- Gene label: chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+ gene_id: chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+
isoforms: [Isoform(gene = chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+, g_start = 0, g_end = 1944, len = 1945,
parts = ['A', 'B', 'C']), Isoform(gene = chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+, g_start = 0, g_end = 1944, len = 1619,
parts = ['A', 'C'])]
- NI, NE, NB: 96, 5, 148
Using default sampler parameters.
Indexing by chromosome...
Output filename ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/chr1/chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+.miso exists, not running MISO.
Submitted total of 2 events.
Loading events from: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/
Writing summary to: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/summary/events_summary.miso
/Library/Frameworks/Python.framework/Versions/2.6/lib/python2.6/site-packages/numpy/lib/npyio.py:1164: DeprecationWarning: The use of `skiprows` is deprecated.
Please use `skip_header` instead.
DeprecationWarning)
- Summarized a total of 2 events.
.Testing gene-level Psi...
Testing GFF indexing of: ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1.mm9.gff
Executing: python ./Projects/tools/MISO/index_gff.py --index ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1.mm9.gff ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed
Indexing GFF...
./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1.mm9.gff appears to already be indexed. Aborting.
Executing: python ./Projects/tools/MISO/run_events_analysis.py --compute-genes-psi ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam --output-dir ./Projects/tools/MISO/test-output/gene-psi-output --read-len 35 --paired-end 250 30
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
Loading settings file from: ./Projects/tools/MISO/settings/miso_settings.txt
Mapping genes to their indexed GFF representation, using ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed
Skipping: ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed/genes_to_filenames.shelve
Computing gene-level Psi for 1 genes...
- GFF index: ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed
- BAM: ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam
- Read length: 35
- Output directory: ./Projects/tools/MISO/test-output/gene-psi-output
- Executing: python ./Projects/tools/MISO/run_miso.py --compute-gene-psi "ENSMUSG00000019943" "./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed/chr10/ENSMUSG00000019943.pickle" ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam ./Projects/tools/MISO/test-output/gene-psi-output --read-len 35 --paired-end 250.0 30.0 --settings-filename ./Projects/tools/MISO/settings/miso_settings.txt
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
Computing Psi for 1 genes...
- ENSMUSG00000019943
- GFF filename: ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed/chr10/ENSMUSG00000019943.pickle
- BAM: ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam
- Outputting to: ./Projects/tools/MISO/test-output/gene-psi-output
- Paired-end mode: (250.0, 30.0)
Loading BAM filename from: ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam
Loading took 0.01 seconds
Aligning reads gene...
Traceback (most recent call last):
File "./Projects/tools/MISO/run_miso.py", line 621, in <module>
main()
File "./Projects/tools/MISO/run_miso.py", line 601, in main
event_type=options.event_type)
File "./Projects/tools/MISO/run_miso.py", line 382, in compute_gene_psi
paired_end=paired_end)
File "./Projects/tools/MISO/sam_utils.py", line 290, in sam_reads_to_isoforms
reads, num_reads = sam_pe_reads_to_isoforms(samfile, gene)
File "./Projects/tools/MISO/sam_utils.py", line 236, in sam_pe_reads_to_isoforms
paired_reads = pair_sam_reads(samfile)
File "./Projects/tools/MISO/sam_utils.py", line 189, in pair_sam_reads
unpaired_reads[read_name] = read
UnboundLocalError: local variable 'read_name' referenced before assignment
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
.
----------------------------------------------------------------------
Ran 3 tests in 4.803s
OK
I am running MISO with paired-end reads but it comes up the error message. Just wondering if anyone had the same problem? Tia!
python test_miso.py
Testing conversion of SAM to BAM...
Executing: python ./Projects/tools/MISO/sam_to_bam.py --convert ./Projects/tools/MISO/test-data/sam-data/c2c12.Atp2b1.sam ./Projects/tools/MISO/test-output/sam-output
Converting SAM to BAM...
- Executing: samtools view -Sbh ./Projects/tools/MISO/test-data/sam-data/c2c12.Atp2b1.sam > ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.bam
[samopen] SAM header is present: 35 sequences.
Sorting BAM file...
- Executing: samtools sort ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.bam ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted
Indexing BAM...
- Executing: samtools index ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam
Conversion took 0.00 minutes.
.Testing single-end SE event interface...
Executing: python ./Projects/tools/MISO/run_events_analysis.py --compute-events-psi se-sample ./Projects/tools/MISO/test-data/se-counts/se_test.counts --output-dir ./Projects/tools/MISO/test-output/SE-output --read-len 35 --overhang-len 4 --event-type SE
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
Loading settings file from: ./Projects/tools/MISO/settings/miso_settings.txt
Computing Psi for events...
Filter events?
- yes
Computing Psi for events of type SE
- samples used: ['se-sample']
Processing sample: label=se-sample, filename=./Projects/tools/MISO/test-data/se-counts/se_test.counts
Filtering events...
Filtering SE events...
Running on a total of 2 events.
Serializing a total of 2 events by Pickle.
Executing: python ./Projects/tools/MISO/run_miso.py --compute-two-iso-psi ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/pickle/se-sample.pickle ./Projects/tools/MISO/test-output/SE-output/SE/se-sample --event-type SE --read-len 35 --overhang-len 4
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
Running MISO on: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/pickle/se-sample.pickle
- Output directory: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/
Called on: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/pickle/se-sample.pickle
Loading from Pickle file took 0.00 seconds.
Parsing of events to genes took 0.00 seconds.
Running sampler on a two-isoform event...
- Gene label: chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:- gene_id: chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:-
isoforms: [Isoform(gene = chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:-, g_start = 0, g_end = 811, len = 812,
parts = ['A', 'B', 'C']), Isoform(gene = chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:-, g_start = 0, g_end = 811, len = 745,
parts = ['A', 'C'])]
- NI, NE, NB: 4, 42, 281
Using default sampler parameters.
Indexing by chromosome...
Output filename ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/chr15/chr15:79605327:79604789:-;chr15:79596283:79596217:-;chr15:79595451:79595246:-.miso exists, not running MISO.
Parsing of events to genes took 0.00 seconds.
Running sampler on a two-isoform event...
- Gene label: chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+ gene_id: chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+
isoforms: [Isoform(gene = chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+, g_start = 0, g_end = 1944, len = 1945,
parts = ['A', 'B', 'C']), Isoform(gene = chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+, g_start = 0, g_end = 1944, len = 1619,
parts = ['A', 'C'])]
- NI, NE, NB: 96, 5, 148
Using default sampler parameters.
Indexing by chromosome...
Output filename ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/chr1/chr1:192752368:192752637:+;chr1:192756746:192757071:+;chr1:192761323:192762671:+.miso exists, not running MISO.
Submitted total of 2 events.
Loading events from: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/
Writing summary to: ./Projects/tools/MISO/test-output/SE-output/SE/se-sample/summary/events_summary.miso
/Library/Frameworks/Python.framework/Versions/2.6/lib/python2.6/site-packages/numpy/lib/npyio.py:1164: DeprecationWarning: The use of `skiprows` is deprecated.
Please use `skip_header` instead.
DeprecationWarning)
- Summarized a total of 2 events.
.Testing gene-level Psi...
Testing GFF indexing of: ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1.mm9.gff
Executing: python ./Projects/tools/MISO/index_gff.py --index ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1.mm9.gff ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed
Indexing GFF...
./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1.mm9.gff appears to already be indexed. Aborting.
Executing: python ./Projects/tools/MISO/run_events_analysis.py --compute-genes-psi ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam --output-dir ./Projects/tools/MISO/test-output/gene-psi-output --read-len 35 --paired-end 250 30
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
Loading settings file from: ./Projects/tools/MISO/settings/miso_settings.txt
Mapping genes to their indexed GFF representation, using ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed
Skipping: ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed/genes_to_filenames.shelve
Computing gene-level Psi for 1 genes...
- GFF index: ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed
- BAM: ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam
- Read length: 35
- Output directory: ./Projects/tools/MISO/test-output/gene-psi-output
- Executing: python ./Projects/tools/MISO/run_miso.py --compute-gene-psi "ENSMUSG00000019943" "./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed/chr10/ENSMUSG00000019943.pickle" ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam ./Projects/tools/MISO/test-output/gene-psi-output --read-len 35 --paired-end 250.0 30.0 --settings-filename ./Projects/tools/MISO/settings/miso_settings.txt
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
Computing Psi for 1 genes...
- ENSMUSG00000019943
- GFF filename: ./Projects/tools/MISO/gff-events/mm9/genes/Atp2b1/indexed/chr10/ENSMUSG00000019943.pickle
- BAM: ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam
- Outputting to: ./Projects/tools/MISO/test-output/gene-psi-output
- Paired-end mode: (250.0, 30.0)
Loading BAM filename from: ./Projects/tools/MISO/test-output/sam-output/c2c12.Atp2b1.sorted.bam
Loading took 0.01 seconds
Aligning reads gene...
Traceback (most recent call last):
File "./Projects/tools/MISO/run_miso.py", line 621, in <module>
main()
File "./Projects/tools/MISO/run_miso.py", line 601, in main
event_type=options.event_type)
File "./Projects/tools/MISO/run_miso.py", line 382, in compute_gene_psi
paired_end=paired_end)
File "./Projects/tools/MISO/sam_utils.py", line 290, in sam_reads_to_isoforms
reads, num_reads = sam_pe_reads_to_isoforms(samfile, gene)
File "./Projects/tools/MISO/sam_utils.py", line 236, in sam_pe_reads_to_isoforms
paired_reads = pair_sam_reads(samfile)
File "./Projects/tools/MISO/sam_utils.py", line 189, in pair_sam_reads
unpaired_reads[read_name] = read
UnboundLocalError: local variable 'read_name' referenced before assignment
Loading settings from: ./Projects/tools/MISO/settings/miso_settings.txt
Settings:
short_queue_name quick
long_queue_name long
cluster_command qsub
filter_results True
min_event_reads 20
lag 10
burn_in 500
num_iters 5000
.
----------------------------------------------------------------------
Ran 3 tests in 4.803s
OK
Comment