ambre_test ERROR workflow.py line 130

[Ilamdri]

Hello,
I tried to run AmBre with the supported example files and afterwards for my own sequences, but I already received an error message for 'workflow.py' which I could not solve by myself after running 'ambre_test':

    ilamdri@watson:~$ ambre_test
    testBlatInstallation (ambre.test.install_unit.TestInstallationValid) ... ok
    testMultiPlxInstallation (ambre.test.install_unit.TestInstallationValid) ... ok
    testPrimer3Installation (ambre.test.install_unit.TestInstallationValid) ... ok
    testTmpDir (ambre.test.install_unit.TestInstallationValid) ... ok
    test_workflow (ambre.test.design_unit.TestDesignWorkflow) ... [-1.0, -0.1, -0.01, -0.001] [10000.0, 100000.0, 1000000.0]
    #Stage1 Primer3Time: 0.0000
    #Number of Primers after Blat: 0
    #Stage2 AlignTime: 0.0001
    #Stage2 MultiPlxTime: 0.0044
    /home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/examples/regions_ex/design_ex.align.out
    /home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/examples/regions_ex/design_ex.multiplx
    /home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/examples/regions_ex/design_ex.multiplx.out
    ERROR
    test_workflow (ambre.test.analyze_unit.TestAnalyze) ... #Stage1 Filtering: 0.0000
    #Stage2 Clustering: 0.0000
    #Stage3 Templating: 0.0063
    ok

    ======================================================================
    ERROR: test_workflow (ambre.test.design_unit.TestDesignWorkflow)
    ----------------------------------------------------------------------
    Traceback (most recent call last):
      File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/test/design_unit.py", line 68, in test_workflow
        pamp_t_bs=(10**4,))
      File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/design/workflow.py", line 344, in run
        self.check_cross_amplification(max_dist=max_cross_amp_dist)
      File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/design/workflow.py", line 130, in check_cross_amplification
        chrs, pos, orient, primer_pos = zip(*alignments)
    ValueError: need more than 0 values to unpack

    ----------------------------------------------------------------------
    Ran 6 tests in 0.013s

    FAILED (errors=1)

Is there anything I could do to solve this error?

Sincerely,
Ilamdri

[l337x911]

It looks like there aren't any off-target alignments, which is why the error is being thrown. Check the blat parameters to make sure you are getting enough off-target hits. Or, check out the zero-cross-align branch ( https://github.com/l337x911/AmBre/tree/zero-cross-aligm) . I haven't tested it, but that may solve your problem.

…

On Tue, Mar 7, 2017 at 4:41 AM, Ilamdri ***@***.***> wrote: Hello, I tried to run AmBre with the supported example files and afterwards for my own sequences, but I already received an error message for 'workflow.py' which I could not solve by myself after running 'ambre_test': ***@***.***:~$ ambre_test testBlatInstallation (ambre.test.install_unit.TestInstallationValid) ... ok testMultiPlxInstallation (ambre.test.install_unit.TestInstallationValid) ... ok testPrimer3Installation (ambre.test.install_unit.TestInstallationValid) ... ok testTmpDir (ambre.test.install_unit.TestInstallationValid) ... ok test_workflow (ambre.test.design_unit.TestDesignWorkflow) ... [-1.0, -0.1, -0.01, -0.001] [10000.0, 100000.0, 1000000.0] #Stage1 Primer3Time: 0.0000 #Number of Primers after Blat: 0 #Stage2 AlignTime: 0.0001 #Stage2 MultiPlxTime: 0.0044 /home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/examples/regions_ex/design_ex.align.out /home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/examples/regions_ex/design_ex.multiplx /home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/examples/regions_ex/design_ex.multiplx.out ERROR test_workflow (ambre.test.analyze_unit.TestAnalyze) ... #Stage1 Filtering: 0.0000 #Stage2 Clustering: 0.0000 #Stage3 Templating: 0.0063 ok ====================================================================== ERROR: test_workflow (ambre.test.design_unit.TestDesignWorkflow) ---------------------------------------------------------------------- Traceback (most recent call last): File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/test/design_unit.py", line 68, in test_workflow pamp_t_bs=(10**4,)) File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/design/workflow.py", line 344, in run self.check_cross_amplification(max_dist=max_cross_amp_dist) File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release-py2.7.egg/ambre/design/workflow.py", line 130, in check_cross_amplification chrs, pos, orient, primer_pos = zip(*alignments) ValueError: need more than 0 values to unpack ---------------------------------------------------------------------- Ran 6 tests in 0.013s FAILED (errors=1) Is there anything I could do to solve this error? Sincerely, Ilamdri — You are receiving this because you are subscribed to this thread. Reply to this email directly, view it on GitHub <#5>, or mute the thread <https://github.com/notifications/unsubscribe-auth/AD2q10zH5CQB7rS7fqbM8Bjs061QBnnyks5rjVBkgaJpZM4MVbkH> .

-- _anand

[Ilamdri]

Thank you for the quick response.

I re-installed AmBre ('zero-cross-align branch') and ran 'ambre_test' again without altering any Blat parameters.

An error for 'regions_ex/design_ex.0000.primer3' was solved by creating the folder 'regions_ex' in the directory 'examples'.
I received empty output-files design_ex.$$$$.primer3.out, design_ex.align.fa, design_ex.align.out, design_ex.multiplx, design_ex.multiplx.out; and
design_ex.$$$$.primer files with content; as well as several empty and a few non-emtpy files in 'default_temp_dir' (*.primer3.out, *.primer3, *.multiplx, *.align.out, *.align.fa)

The previously mentioned error ('ValueError: need more than 0 values to unpack') was skipped, but a new error occured. Could it be that it does not design any primers?
Should it not design primers from the provided files in the 'examples' directory (reference.fasta, regions.test)?

I do not know if this might be relevant, but some of the provided scripts contain hard-coded lines:

/design/parse_primer3.py 	line 169-172, line 187
/design/summary_links.py 	line 255
/scripts/convert_mrna.py 	line 8-9
/scripts/get_fa_from_input.py 	line 3-4
/scripts/get_fasta_seq.py 	line 5
/scripts/plot_bp.py 		line 146
/scripts/plot_pop_del.py 	line 6-7
/utils/reference.py 		line 112
/utils/reference.py 		line 115
/utils/SAM.py 			line 109-114

ilamdri@watson:~$ ambre_test
testBlatInstallation (ambre.test.install_unit.TestInstallationValid) ... ok
testMultiPlxInstallation (ambre.test.install_unit.TestInstallationValid) ... ok
testPrimer3Installation (ambre.test.install_unit.TestInstallationValid) ... ok
testTmpDir (ambre.test.install_unit.TestInstallationValid) ... ok
test_workflow (ambre.test.design_unit.TestDesignWorkflow) ... [-1.0, -0.1, -0.01, -0.001] [10000.0, 100000.0, 1000000.0]
#Stage1 Primer3Time: 0.0001
#Number of Primers after Blat: 0
#Stage2 AlignTime: 0.0000
#Stage2 MultiPlxTime: 0.0000
#Number of Cross Amplifications 0
#Stage2 CrossAmpTime: 0.0000
ERROR: No result!!!
#Region 0, 0-10000 has 0 primers
Error Region 0, 0-10000 has no primers to choose from
/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/examples/regions_ex/design_ex.align.out
/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/examples/regions_ex/design_ex.multiplx.out
ERROR
test_workflow (ambre.test.analyze_unit.TestAnalyze) ... #Stage1 Filtering: 0.0000
#Stage2 Clustering: 0.0000
#Stage3 Templating: 0.0064
ok

======================================================================
ERROR: test_workflow (ambre.test.design_unit.TestDesignWorkflow)
----------------------------------------------------------------------
Traceback (most recent call last):
  File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/test/design_unit.py", line 67, in test_workflow
    pamp_t_bs=(10**4,))
  File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/design/workflow.py", line 404, in run
    primer_graph = sa_cost.get_primer_graph(multiplx_out_fpath, regions_pamp, d=self.d, rho=self.rho)
  File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/design/sa_cost.py", line 490, in get_primer_graph
    graph.load(multiplx_filter_func)
  File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/design/sa_cost.py", line 130, in load
    self.load_primer_maps()
  File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/design/sa_cost.py", line 120, in load
    f.next()
StopIteration

----------------------------------------------------------------------
Ran 6 tests in 0.008s

FAILED (errors=1)

Sincerely,
Ilamdri

[l337x911]

Based on your log, it looks like primer3 is not generating primers. Please make sure the primer3 is configured correctly.

…

On Mar 10, 2017 2:33 AM, "Ilamdri" ***@***.***> wrote: Thank you for the quick response. I re-installed AmBre ('zero-cross-align branch') and ran 'ambre_test' again without altering any Blat parameters. An error for 'regions_ex/design_ex.0000.primer3' was solved by creating the folder 'regions_ex' in the directory 'examples'. I received empty output-files design_ex.$$$$.primer3.out, design_ex.align.fa, design_ex.align.out, design_ex.multiplx, design_ex.multiplx.out; and design_ex.$$$$.primer files with content; as well as several empty and a few non-emtpy files in 'default_temp_dir' (*.primer3.out, *.primer3, *.multiplx, *.align.out, *.align.fa) The previously mentioned error ('ValueError: need more than 0 values to unpack') was skipped, but a new error occured. Could it be that it does not design any primers? Should it not design primers from the provided files in the 'examples' directory (reference.fasta, regions.test)? I do not know if this might be relevant, but some of the provided scripts contain hard-coded lines: /design/parse_primer3.py line 169-172, line 187 /design/summary_links.py line 255 /scripts/convert_mrna.py line 8-9 /scripts/get_fa_from_input.py line 3-4 /scripts/get_fasta_seq.py line 5 /scripts/plot_bp.py line 146 /scripts/plot_pop_del.py line 6-7 /utils/reference.py line 112 /utils/reference.py line 115 /utils/SAM.py line 109-114 ***@***.***:~$ ambre_test testBlatInstallation (ambre.test.install_unit.TestInstallationValid) ... ok testMultiPlxInstallation (ambre.test.install_unit.TestInstallationValid) ... ok testPrimer3Installation (ambre.test.install_unit.TestInstallationValid) ... ok testTmpDir (ambre.test.install_unit.TestInstallationValid) ... ok test_workflow (ambre.test.design_unit.TestDesignWorkflow) ... [-1.0, -0.1, -0.01, -0.001] [10000.0, 100000.0, 1000000.0] #Stage1 Primer3Time: 0.0001 #Number of Primers after Blat: 0 #Stage2 AlignTime: 0.0000 #Stage2 MultiPlxTime: 0.0000 #Number of Cross Amplifications 0 #Stage2 CrossAmpTime: 0.0000 ERROR: No result!!! #Region 0, 0-10000 has 0 primers Error Region 0, 0-10000 has no primers to choose from /home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/examples/regions_ex/design_ex.align.out /home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/examples/regions_ex/design_ex.multiplx.out ERROR test_workflow (ambre.test.analyze_unit.TestAnalyze) ... #Stage1 Filtering: 0.0000 #Stage2 Clustering: 0.0000 #Stage3 Templating: 0.0064 ok ====================================================================== ERROR: test_workflow (ambre.test.design_unit.TestDesignWorkflow) ---------------------------------------------------------------------- Traceback (most recent call last): File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/test/design_unit.py", line 67, in test_workflow pamp_t_bs=(10**4,)) File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/design/workflow.py", line 404, in run primer_graph = sa_cost.get_primer_graph(multiplx_out_fpath, regions_pamp, d=self.d, rho=self.rho) File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/design/sa_cost.py", line 490, in get_primer_graph graph.load(multiplx_filter_func) File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/design/sa_cost.py", line 130, in load self.load_primer_maps() File "/home/ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/design/sa_cost.py", line 120, in load f.next() StopIteration ---------------------------------------------------------------------- Ran 6 tests in 0.008s FAILED (errors=1) Sincerely, Ilamdri — You are receiving this because you commented. Reply to this email directly, view it on GitHub <#5 (comment)>, or mute the thread <https://github.com/notifications/unsubscribe-auth/AD2q10GDlG17alTg9bZ9sTZ1x8gicntHks5rkScEgaJpZM4MVbkH> .

[Ilamdri]

I had no problems generating primers with primer3_core. Then I tried to run 'ambre_design' on a reference sequence-file and a regions-file formatted like the AmBre example files 'reference.fasta' and 'regions.test'. The same error as in 'ambre_test' occured, but I also received some file output ('$$$$.0001.primer3', '$$$$.0001.primer3.out') and a message primer3 output: None.

I added the PRIMER_THERMODYNAMIC_PARAMETERS_PATH= in the '$$$$.0001.primer3' file, ran primer3_core, but did not receive any primers either. But when I deleted the line SEQUENCE_PRIMER_PAIR_OK_REGION_LIST=0,5,,, primer3_core generated all 75 primer-pairs as defined in '$$$$.0001.primer3'.

Might it be possible that the error is caused by this string?

Sincerely,
Ilamdri

[Ilamdri]

I now somehow circumvented the error by setting line 76 in 'workflow.py' (print >>primer3_in, "SEQUENCE_PRIMER_PAIR_OK_REGION_LIST=%s"%(ok_reg)) as comment.

Running 'ambre_test' resulted in a 'design_ex.out'-file, containing two sets of PAMP primers.

Maybe the error is caused somewhere at line 65 in 'workflow.py':

  def _prepare_input(self, fpath, seq, forward_flag):
    if forward_flag:
      ok_reg = ",,%d,5"%(len(seq)-5)
    else:
      ok_reg = "0,5,,"

Ilamdri@watson:~$ ambre_test
testBlatInstallation (ambre.test.install_unit.TestInstallationValid) ... ok
testMultiPlxInstallation (ambre.test.install_unit.TestInstallationValid) ... ok
testPrimer3Installation (ambre.test.install_unit.TestInstallationValid) ... ok
testTmpDir (ambre.test.install_unit.TestInstallationValid) ... ok
test_workflow (ambre.test.design_unit.TestDesignWorkflow) ... [-1.0, -0.1, -0.01, -0.001] [10000.0, 100000.0, 1000000.0]

primer3 output: None
#Stage1 Primer3Time: 7.5654

#Number of Primers after Blat: 211
#Stage2 AlignTime: 0.8102
#Stage2 MultiPlxTime: 0.0039
#Number of Cross Amplifications 3428
#Stage2 CrossAmpTime: 0.0297
#Region 0, 0-10000 has 74 primers
#Region 1, 10000-20000 has 137 primers
18000.0 ['8478', '13721']
18000.0 ['8161', '11998']
#Stage3 PAMPTime: 117.4844

ok
test_workflow (ambre.test.analyze_unit.TestAnalyze) ... #Stage1 Filtering: 1.7387
Data:  (794, 3)
UniqueData:  (696, 3)
Elements 696
SweepLines 225
ClusterCount: 15
Size:81, -17461, -92452, 2
Size:79, 92127, 308317, 0
Size:78, -92454, -17463, 2
Size:77, 308317, 92127, 0
Size:53, -50641, 177515, 6
Size:53, -12687, 177809, 17
Size:52, 174622, -4534, 4
Size:52, 177515, -50641, 16
Size:52, 177809, -12687, 5
Size:51, -155806, 170230, 4
Size:51, -4534, 174622, 25
Size:50, 170230, -155806, 4
#Stage2 Clustering: 0.0455
#Stage3 Templating: 0.0037
ok

----------------------------------------------------------------------
Ran 6 tests in 127.724s

OK

Sincerely,
Ilamdri

[Ilamdri]

I now ran 'ambre_design', but all output sequences (either if t<is_forward> is True or False) are in reverse orientation. Is it supposed to do so, or might it be caused by my deactivation of line 76 in 'workflow.py'?

Input:
deleted region from MTAP ex8 to ANRIL ex1 in the provided 'reference.fasta'-file

'regions.test'
MTAP 44300 53499 forward
MTAP 151000 53499 reverse

deactivated 'workflow.py' line 76 (print >>primer3_in, "SEQUENCE_PRIMER_PAIR_OK_REGION_LIST=%s"%(ok_reg))

Output:

Ilamdri@watson:~/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/examples$ ambre_design reference.fasta regions.test
/home/Ilamdri/.local/bin/primer3/src/primer3_core -p3_settings_file=/home/Ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/data/primer3_A01.txt /home/Ilamdri/temp/hW4qTZ.0000.primer3
/home/Ilamdri/.local/bin/primer3/src/primer3_core -p3_settings_file=/home/Ilamdri/.local/lib/python2.7/site-packages/AmBre-1.0release_r0-py2.7.egg/ambre/data/primer3_A01.txt /home/Ilamdri/temp/hW4qTZ.0001.primer3
primer3 output: None
#Stage1 Primer3Time: 27.0825
8644 /home/Ilamdri/.linuxbrew/bin/blat -t=dna -q=dna -stepSize=2 -tileSize=8 -repMatch=1048576 -minScore=15 -noHead reference.fasta /home/Ilamdri/temp/hW4qTZ.align.fa /home/Ilamdri/temp/hW4qTZ.align.out
#Number of Primers after Blat: 571
#Stage2 AlignTime: 2.0579
#Stage2 MultiPlxTime: 0.0071
#Number of Cross Amplifications 14835
#Stage2 CrossAmpTime: 0.1345
#Region 0, 0-53499 has 241 primers
#Region 1, 53499-106998 has 330 primers
93998.0 ['45686', '89015']
93998.0 ['40319', '88517']
93998.0 ['30969', '100507']
93998.0 ['30973', '83607']
93998.0 ['12988', '83350']
93998.0 ['40109', '88514']
98718.0 ['40129', '56989']
99266.0 ['20991', '56715']
93998.0 ['5397', '83424']
93998.0 ['5428', '102941']
99216.0 ['31024', '56740']
93998.0 ['5465', '88514']
93998.0 ['5365', '90781']
93998.0 ['40294', '78701']
93998.0 ['4155', '103323']
93998.0 ['30958', '83198']
93998.0 ['20991', '86634']
93998.0 ['30998', '98457']
93998.0 ['30940', '72046']
93998.0 ['36552', '83418']
93998.0 ['31234', '100551']
93998.0 ['44937', '80064']
93998.0 ['12982', '103301']
93998.0 ['30960', '100492']
93998.0 ['30934', '83239']
93998.0 ['5432', '83220']
93998.0 ['40131', '100510']
93998.0 ['14178', '86445']
93998.0 ['40113', '103297']
99210.0 ['36455', '56743']
93998.0 ['5302', '88487']
99264.0 ['36232', '56716']
93998.0 ['4019', '103297']
93998.0 ['1718', '100492']
93998.0 ['40138', '76308']
93998.0 ['30985', '100485']
#Stage3 PAMPTime: 18039.1806
/home/Ilamdri/temp/hW4qTZ.0001.primer3.out
/home/Ilamdri/temp/hW4qTZ.align.fa
/home/Ilamdri/temp/hW4qTZ.align.out
/home/Ilamdri/temp/hW4qTZ.multiplx
/home/Ilamdri/temp/hW4qTZ.multiplx.out
/home/Ilamdri/temp/hW4qTZ.sa
14339	894.04	-03,04,00
MTAP	46015	True	ggccaaagcgggtggatcacgagg	1.1792
MTAP	49720	True	TCCTGAGGGTGCCACCGTACCAGAG	0.8613
MTAP	55935	True	ttagctgggcatggtggcaggcacc	1.1207
MTAP	60961	True	gtgggaagcggtgtggggattcctc	1.0579
MTAP	65294	True	atggcactggcatctggtgagggcc	0.6897
MTAP	70600	True	tttggggactcactggggcagggtg	0.0414
MTAP	181109	False	ggcagcttccacgtggtgttgaccc	0.0072
MTAP	80491	True	gctgcacccactgtctgacaagccc	0.1993
MTAP	179140	False	ccacccgccttggcctctcaaactg	0.4042
MTAP	188937	False	ccctgctatgtggccagctctgtgc	0.5225
MTAP	157196	False	TCAGGCCCTGTCCTGTGGCATCTCC	0.3267
MTAP	169549	False	TCCCTTTCAGAGCCTGCCCCAGCTG	0.1275
MTAP	176312	False	aaaaatCAGGCAggctgggcgtggtg	1.0874
MTAP	181303	False	ttcatggcagcccctcccatcacagg	1.0505
MTAP	186442	False	CTGGACGGGAGAAGTGCTGACGGAGG	1.0167
MTAP	192001	False	GCTGTGCACCAGAGGCAGGCATTTG	1.1048
MTAP	197524	False	ttcaagaccagccgggccaacatgg	0.6538
MTAP	203227	False	TGGTGGTTGAAGCAAGACCCCAGCC	0.9734
=
14362	9023.24	-03,05,02
MTAP	46017	True	gaggccaaagcgggtggatcacgag	0.5505
MTAP	49732	True	ACTGTCCTCCACTCCTGAGGGTGCC	0.8919
MTAP	200822	False	CACCCAGGACAGGACCCAGCACACC	0.5425
MTAP	60961	True	gtgggaagcggtgtggggattcctc	1.0579
MTAP	201045	False	ATGCCGGTGGAGAAGGGACATGGGC	0.4559
MTAP	70600	True	tttggggactcactggggcagggtg	0.0414
MTAP	75778	True	tgcccaaagtgtcagggggaaggtcc	1.1065
MTAP	80855	True	aggcaagcctccttgagctgtggtg	1.1583
MTAP	179140	False	ccacccgccttggcctctcaaactg	0.4042
MTAP	188937	False	ccctgctatgtggccagctctgtgc	0.5225
MTAP	157196	False	TCAGGCCCTGTCCTGTGGCATCTCC	0.3267
MTAP	169549	False	TCCCTTTCAGAGCCTGCCCCAGCTG	0.1275
MTAP	176312	False	aaaaatCAGGCAggctgggcgtggtg	1.0874
MTAP	181303	False	ttcatggcagcccctcccatcacagg	1.0505
MTAP	186516	False	TGGAAAGAGGAACCTGGTGGGTGGCC	1.1065
MTAP	192001	False	GCTGTGCACCAGAGGCAGGCATTTG	1.1048
MTAP	197532	False	gtcaggagttcaagaccagccgggc	0.8629
MTAP	203227	False	TGGTGGTTGAAGCAAGACCCCAGCC	0.9734
=
14736	8998.59	-03,05,00
MTAP	181302	False	tcatggcagcccctcccatcacagg	0.0229
MTAP	179429	False	gctcttgctgcccaggctgaagtgc	0.3364
MTAP	55936	True	attagctgggcatggtggcaggcac	0.5310
MTAP	60968	True	aaccattgtgggaagcggtgtgggg	0.6951
MTAP	65294	True	atggcactggcatctggtgagggcc	0.6897
MTAP	70600	True	tttggggactcactggggcagggtg	0.0414
MTAP	179429	False	gctcttgctgcccaggctgaagtgc	0.3364
MTAP	80782	True	cctcagcaatggcagacacccctcc	1.0480
MTAP	179140	False	ccacccgccttggcctctcaaactg	0.4042
MTAP	188937	False	ccctgctatgtggccagctctgtgc	0.5225
MTAP	157196	False	TCAGGCCCTGTCCTGTGGCATCTCC	0.3267
MTAP	169551	False	TCTCCCTTTCAGAGCCTGCCCCAGC	0.0985
MTAP	173817	False	GTCCACATGGGCAGCCTGGGTCTTC	0.8284
MTAP	179142	False	atccacccgccttggcctctcaaac	1.0969
MTAP	185097	False	CCCAGCTGCCTGTCCTCCAGTGCTC	0.6234
MTAP	191208	False	aacctgggaggcggaggtagcagtg	0.3500
MTAP	197004	False	TCCCACCTCCTTTGCTGTTGCACCC	0.9734
MTAP	203227	False	TGGTGGTTGAAGCAAGACCCCAGCC	0.9734
=
14865	8777.70	-03,05,01
MTAP	46065	True	AAACTTggctgggcgcagtggctc	1.1888
MTAP	198038	False	agcctgggaggtggaggttgcagtg	0.0695
MTAP	180875	False	agggccatcgtccttcagaccccag	0.5534
MTAP	60961	True	gtgggaagcggtgtggggattcctc	1.0579
MTAP	65294	True	atggcactggcatctggtgagggcc	0.6897
MTAP	70600	True	tttggggactcactggggcagggtg	0.0414
MTAP	179429	False	gctcttgctgcccaggctgaagtgc	0.3364
MTAP	80686	True	tgggaccctctgagctaggcatggg	0.9903
MTAP	179140	False	ccacccgccttggcctctcaaactg	0.4042
MTAP	188937	False	ccctgctatgtggccagctctgtgc	0.5225
MTAP	157196	False	TCAGGCCCTGTCCTGTGGCATCTCC	0.3267
MTAP	169549	False	TCCCTTTCAGAGCCTGCCCCAGCTG	0.1275
MTAP	176312	False	aaaaatCAGGCAggctgggcgtggtg	1.0874
MTAP	181303	False	ttcatggcagcccctcccatcacagg	1.0505
MTAP	186504	False	CCTGGTGGGTGGCCTAGATGGGTGTC	1.1916
MTAP	192616	False	aagatcgcgcaactgcactccagcc	0.1291
MTAP	198020	False	tgcagtgagccaagattgcgccactg	1.0871
MTAP	203227	False	TGGTGGTTGAAGCAAGACCCCAGCC	0.9734
=
15473	900.80	-03,04,01
MTAP	181302	False	tcatggcagcccctcccatcacagg	0.0229
MTAP	198038	False	agcctgggaggtggaggttgcagtg	0.0695
MTAP	200822	False	CACCCAGGACAGGACCCAGCACACC	0.5425
MTAP	60968	True	aaccattgtgggaagcggtgtgggg	0.6951
MTAP	65294	True	atggcactggcatctggtgagggcc	0.6897
MTAP	70600	True	tttggggactcactggggcagggtg	0.0414
MTAP	188803	False	gtgtgtgtgtgcgtgcgtgtgtgtg	0.3982
MTAP	198040	False	tgagcctgggaggtggaggttgcag	0.1584
MTAP	178994	False	caacctccacctcgcaggctcaagc	0.3805
MTAP	188937	False	ccctgctatgtggccagctctgtgc	0.5225
MTAP	157196	False	TCAGGCCCTGTCCTGTGGCATCTCC	0.3267
MTAP	169549	False	TCCCTTTCAGAGCCTGCCCCAGCTG	0.1275
MTAP	176106	False	tctcctgcctctgcctcccaaggag	1.0573
MTAP	181084	False	agggtcaacaccacgtggaagctgc	0.7408
MTAP	186416	False	ACCTCCGTCAGCACTTCTCCCGTCC	0.3904
MTAP	191822	False	aaggatagggcgtcctgggcagagg	0.7224
MTAP	198066	False	gagtagctgggactacaggcacgggc	1.0507
MTAP	202748	False	ACTCCAGAGACTTGGCCCTGCCTGG	0.3776
=

Sincerely,
Ilamdri