[ensembl-dev] VEP error: Forked process failed.

Will McLaren wm2 at ebi.ac.uk
Tue May 14 11:22:44 BST 2013


Thanks - try
http://cvs.sanger.ac.uk/cgi-bin/viewvc.cgi/ensembl-variation/modules/Bio/EnsEMBL/Variation/Utils/VEP.pm?revision=1.93&root=ensembl

Will


On 14 May 2013 11:09, Duarte Molha <duartemolha at gmail.com> wrote:

> It seems the problems are still there :
>
> Here is my output:
>
> perl variant_effect_predictor.pl --config vep_human.ini -i INPUT.vcf
> --fork 16
>
> 2013-05-14 10:33:38 - Read configuration from vep_human.ini
> #----------------------------------#
> # ENSEMBL VARIANT EFFECT PREDICTOR #
> #----------------------------------#
>
> version 71
>
> By Will McLaren (wm2 at ebi.ac.uk)
>
> Configuration options:
>
> ###
> allow_non_variant    1
> buffer_size                 500000
> cache                1
> canonical            1
> ccds                 1
> check_alleles        1
> check_existing       1
> config               vep_human.ini
> core_type            core
> custom
> /ReferenceData/vep_additional_annotations/Somatic_variation_phenotypes.bed.gz,Somatic,bed,exact
>
>
> /ReferenceData/vep_additional_annotations/dbsnp135_ensembl_variation_phenotype.bed.gz,dbsnp135,bed,exact
> dir                       /ReferenceData/vep_cache/
> domains                 1
> force_overwrite      1
> fork                          16
> gmaf                 1
> hgnc                 1
> host                 ensembldb.ensembl.org
> individual           all
> input_file           INPUT.vcf
> numbers              1
> plugin               Blosum62
> Condel,/ReferenceData/vep_cache/Plugins/config/Condel/config,b  Carol
> polyphen             b
> port                 5306
> protein              1
> regulatory           1
> sift                 b
> species              homo_sapiens
> stats                HASH(0x35a8000)
> terms                SO
> toplevel_dir         /ReferenceData/vep_cache/
> verbose              1
> xref_refseq          1
>
> --------------------
>
> Will only load v71 databases
> Species 'homo_sapiens' loaded from database 'homo_sapiens_core_71_37'
> Species 'homo_sapiens' loaded from database 'homo_sapiens_cdna_71_37'
> Species 'homo_sapiens' loaded from database 'homo_sapiens_vega_71_37'
> Species 'homo_sapiens' loaded from database
> 'homo_sapiens_otherfeatures_71_37'
> Species 'homo_sapiens' loaded from database 'homo_sapiens_rnaseq_71_37'
> homo_sapiens_variation_71_37 loaded
> homo_sapiens_funcgen_71_37 loaded
> Bio::EnsEMBL::Compara::DBSQL::DBAdaptor not found so the following compara
> databases will be ignored: ensembl_compara_71
> ensembl_ancestral_71 loaded
> ensembl_ontology_71 loaded
> 2013-05-14 10:33:39 - Connected to core version 71 database and variation
> version 71 database
> 2013-05-14 10:33:39 - Read existing cache info
> 2013-05-14 10:33:39 - Loaded plugin: Blosum62
> 2013-05-14 10:33:39 - Loaded plugin: Condel
> 2013-05-14 10:33:39 - Loaded plugin: Carol
> 2013-05-14 10:33:40 - Starting...
> 2013-05-14 10:33:40 - Detected format of input file as vcf
> 2013-05-14 10:33:46 - Read 195789 variants into buffer
> 2013-05-14 10:33:46 - Skipping 67552 non-variant loci
> 2013-05-14 10:33:46 - Reading transcript data from cache and/or database
> [======================================================================================================]
>  [ 100% ]
> 2013-05-14 10:40:19 - Retrieved 189344 transcripts (0 mem, 202901 cached,
> 0 DB, 13557 duplicates)
> 2013-05-14 10:40:19 - Reading regulatory data from cache and/or database
> [======================================================================================================]
>  [ 100% ]
> 2013-05-14 10:50:09 - Retrieved 872092 regulatory features (0 mem, 872351
> cached, 0 DB, 259 duplicates)
> 2013-05-14 10:50:12 - Calculating consequences
> Use of uninitialized value $_ in pattern match (m//) at
> /NGS_Test/duarte/vep_71/Bio/EnsEMBL/Variation/Utils/VEP.pm line 1022.
> Use of uninitialized value $_ in pattern match (m//) at
> /NGS_Test/duarte/vep_71/Bio/EnsEMBL/Variation/Utils/VEP.pm line 1030.
> Use of uninitialized value $_ in pattern match (m//) at
> /NGS_Test/duarte/vep_71/Bio/EnsEMBL/Variation/Utils/VEP.pm line 1037.
> Use of uninitialized value $_ in pattern match (m//) at
> /NGS_Test/duarte/vep_71/Bio/EnsEMBL/Variation/Utils/VEP.pm line 1089.
> Use of uninitialized value $_ in concatenation (.) or string at
> /NGS_Test/duarte/vep_71/Bio/EnsEMBL/Variation/Utils/VEP.pm line 1097.
>
> ERROR: Forked process failed
>
>
>
>
> =========================
>      Duarte Miguel Paulo Molha
>          http://about.me/duarte
> =========================
>
>
> On Tue, May 14, 2013 at 10:42 AM, Duarte Molha <duartemolha at gmail.com>wrote:
>
>> Thanks
>>
>> Running a annotation using 16 forks... lets see how it handles :)
>> I'll report back any issues.
>>
>> Thanks for the update
>>
>> Duarte
>>
>>
>> =========================
>>      Duarte Miguel Paulo Molha
>>          http://about.me/duarte
>> =========================
>>
>>
>> On Tue, May 14, 2013 at 10:16 AM, Will McLaren <wm2 at ebi.ac.uk> wrote:
>>
>>> Stuart, Guillermo, Duarte,
>>>
>>> I'm currently working on some code as I stated above to improve
>>> stability and performance under forking.
>>>
>>> I've committed some code to the HEAD of our CVS tree which should help
>>> the problems you are encountering. You'd all be welcome to test this out,
>>> with the obvious proviso that this is development code and may contain bugs!
>>>
>>> To use this, you should download the copy of VEP.pm from:
>>>
>>>
>>> http://cvs.sanger.ac.uk/cgi-bin/viewvc.cgi/ensembl-variation/modules/Bio/EnsEMBL/Variation/Utils/VEP.pm?revision=1.92&root=ensembl
>>>
>>> and replace the VEP.pm under
>>> ensembl-variation/modules/Bio/EnsEMBL/Variation/Utils (or just
>>> Bio/EnsEMBL/Variation/Utils if you use INSTALL.pl) with this one.
>>>
>>> This code will appear in production in the next proper release of
>>> Ensembl.
>>>
>>> Regards
>>>
>>> Will
>>>
>>>
>>> On 14 May 2013 09:55, Stuart Meacham <sm766 at cam.ac.uk> wrote:
>>>
>>>>  Hi,
>>>>
>>>> I certainly don't want to hijack this thread but it seemed daft to
>>>> start another. I am also getting forking errors. I don't use any custom
>>>> plugins and am using a validated VCF as input (with about 600,000
>>>> variants). Trying to fork more than 4 threads is unstable even on my
>>>> machine which has 64 cores and half a TB of RAM.
>>>>
>>>> I haven't found anything reproducible, however if I do I'll report back
>>>> to the list.
>>>>
>>>> Thanks
>>>>
>>>> Stuart
>>>>
>>>>
>>>> On 14/05/2013 09:42, Will McLaren wrote:
>>>>
>>>> Hello,
>>>>
>>>>  Your aa_grantham_distance plugin is somewhat inefficient - it
>>>> retrieves the peptide alleles from the HGVS annotation, which itself
>>>> requires some database fetching and processing to produce. This is why it
>>>> is slow.
>>>>
>>>>  You can get the peptides from the transcript variation object:
>>>>
>>>>  my @peps = split "/", $tva->transcript_variation->pep_allele_string();
>>>>
>>>>  This will give you single-letter AA codes, but you could either
>>>> modify your hash or use BioPerl to convert:
>>>>
>>>>  $seqobj = Bio::PrimarySeq->new ( -seq => $single_letter_aa);
>>>> $three_letter_aa = Bio::SeqUtils->seq3($seqobj);
>>>>
>>>>  You should also declare your distances hash in the new() sub and
>>>> store it on $self; this will also marginally speed up your plugin.
>>>>
>>>>  Regarding the forking issues, we are working on improving stability
>>>> under forking.
>>>>
>>>>  Thanks for your patience
>>>>
>>>>  Will
>>>>
>>>>
>>>>  On 14 May 2013 07:37, Guillermo Marco Puche <
>>>> guillermo.marco at sistemasgenomicos.com> wrote:
>>>>
>>>>>  Hello,
>>>>>
>>>>> I'm not really sure which one of those plugins is causing the fork
>>>>> error. I cannot recreate it now running each one of them separately.
>>>>>
>>>>> Here are both:
>>>>>
>>>>> https://github.com/guillermomarco/vep_plugins_71
>>>>>
>>>>> They also slow the calculating consequences process a lot.
>>>>> aa_grantham_distance.pm is just a hardcoded plugin from one of the
>>>>> biologists in my work. It was just a pure copy paste and adaptation to make
>>>>> it work as a VEP plugin. Maybe the problem is in the matrix definition
>>>>> every time the sub routine is called. I'm not running out of memory nor
>>>>> CPU. I'm currently using it with 2 threads and buffersize of 500 for a 5000
>>>>> variant vcf file.
>>>>>
>>>>> I'm my honest opinion, I think one (or even both) of those plugins are
>>>>> slowing so much the calculating process that sometimes the fork just dies.
>>>>> Like when you have a timeout during to heavy network traffic. So when you
>>>>> use them together with lot of other plugins like Condel, Consequence, etc..
>>>>> they may be causing the process to handle and die.
>>>>>
>>>>> Best regards,
>>>>> Guillermo.
>>>>>
>>>>>
>>>>> On 05/13/2013 03:55 PM, Duarte Molha wrote:
>>>>>
>>>>> I also get this error... it is so prevalent and so difficult to
>>>>> pinpoint what is causing it that I have given up on forking my annotation
>>>>> process.
>>>>>
>>>>>  I do think it is related to the number of forks. It seems to crash
>>>>> less often if you use a low number of forks... anything above 5
>>>>> will undoubtedly crash the script at least in my experience.
>>>>>
>>>>>  Cheers
>>>>>
>>>>> Duarte
>>>>>
>>>>> =========================
>>>>>      Duarte Miguel Paulo Molha
>>>>>           http://about.me/duarte
>>>>> =========================
>>>>>
>>>>>
>>>>> On Mon, May 13, 2013 at 2:50 PM, Will McLaren <wm2 at ebi.ac.uk> wrote:
>>>>>
>>>>>> Hi Guillermo,
>>>>>>
>>>>>>  Test each plugin individually until you find the one that causes
>>>>>> the error. It is highly unlikely that a particular combination of plugins
>>>>>> is causing the crash.
>>>>>>
>>>>>>  Check that there are no "print" (to STDOUT or STDERR) statements in
>>>>>> your plugin - forking assumes that code remains silent otherwise it will
>>>>>> throw errors like this.
>>>>>>
>>>>>>  Also, check what, if anything, is cached between runs of your
>>>>>> plugin. If you are caching things (for example to avoid re-querying a
>>>>>> database), you may need to write storable hooks to ensure the data is
>>>>>> getting cached between forks - see
>>>>>> https://github.com/ensembl-variation/VEP_plugins/blob/master/ProteinSeqs.pmfor an example.
>>>>>>
>>>>>>  If you still have no luck, send me the code and an input file that
>>>>>> recreates the problem.
>>>>>>
>>>>>>  Regards
>>>>>>
>>>>>>  Will
>>>>>>
>>>>>>
>>>>>>  On 13 May 2013 13:18, Guillermo Marco Puche <
>>>>>> guillermo.marco at sistemasgenomicos.com> wrote:
>>>>>>
>>>>>>>   Hello,
>>>>>>>
>>>>>>> I've started to recently having problems with VEP script while using
>>>>>>> different plugins (most of them own plugins).
>>>>>>>
>>>>>>> 2013-05-13 13:59:44 - Connected to core version 71 database and variation version 71 database
>>>>>>> 2013-05-13 13:59:44 - Loaded plugin: vcf_input
>>>>>>> 2013-05-13 13:59:44 - Loaded plugin: biobase
>>>>>>> 2013-05-13 13:59:44 - Loaded plugin: aa_grantham_distance
>>>>>>> 2013-05-13 13:59:44 - Loaded plugin: flanking_sequence
>>>>>>> 2013-05-13 13:59:44 - Loaded plugin: Condel
>>>>>>> 2013-05-13 13:59:44 - Output fields redefined (37 defined)
>>>>>>> 2013-05-13 13:59:44 - Starting...
>>>>>>> 2013-05-13 13:59:45 - Read 3888 variants into buffer
>>>>>>> 2013-05-13 13:59:54 - Reading transcript data from cache and/or database
>>>>>>> [===============================================]  [ 100% ]
>>>>>>> 2013-05-13 14:02:38 - Retrieved 6463 transcripts (0 mem, 0 cached, 13743 DB, 7280 duplicates)
>>>>>>> 2013-05-13 14:02:38 - Calculating consequences
>>>>>>> [===================================>           ]   [ 78% ]
>>>>>>> ERROR: Forked process failed
>>>>>>>
>>>>>>>
>>>>>>>
>>>>>>> I'm not getting any other error message. So I cannot debug properly.
>>>>>>> I thought my plugins were OK but it's seems they don't. I think the problem
>>>>>>> occurs when I use "aa_grantham_distance plugin" together with
>>>>>>> "flanking_sequence". I've no idea what could be causing this.
>>>>>>>
>>>>>>> I'm running VEP on verbose mode but I can't get any usefull
>>>>>>> information. How could I debug that?
>>>>>>>
>>>>>>> Best regards,
>>>>>>> Guillermo.
>>>>>>>
>>>>>>>
>>>>>>>  _______________________________________________
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>>>>>>>
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>>>>
>>>>
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