[maker-devel] Maker not producing expected output

Daniel Ence dence at genetics.utah.edu
Sat Oct 17 09:46:09 MDT 2015


Hi Kevin, So I have a couple of clarifying questions, and an explanation that’ll hopefully be helpful. 

If you look in the master datastore log, do you see an entry that shows that scaffold finished successfully? It will have the name of the scaffold, then the path to the results directory, and then a status. There should be one that shows that maker started working on it, and one that shows that maker finished it. 

Second what are the files that you’re expecting to see? I think you’re expecting to see couple of fasta files and a gff3 file that contain all the annotation results all gathered together. You can gather those results with the fasta_merge, and gff3_merge scripts that came with maker. 

To explain what you saw in that example results directory that you sent, if there weren’t any models or predictions on that scaffold, then there won’t be fasta files in the results directory. You could verify that by looking at the scaffold-334630.gff file. The fast_merge, and gff3_merge will gather all of the results fasta and gff files for all the scaffolds and put them into a few fasta files and one gff3 files, respectively. 

Let me know whether that helps, 
Daniel


Daniel Ence
Graduate Student
Eccles Institute of Human Genetics
University of Utah
15 North 2030 East, Room 2100
Salt Lake City, UT 84112-5330

> On Oct 16, 2015, at 10:10 PM, Kevin Kocot <kmkocot at gmail.com> wrote:
> 
> Hello,
> 
> I've run Maker on a draft invertebrate genome and it seemed to finish successfully. However, many of the expected output files were not produced. If I go to, for example, XX_datastore/00/0C/scaffold-334630/, all I see is:
> 
> theVoid.scaffold-334630
> run.log
> scaffold-334630.gff
> 
> In particular, I'm looking for the transcripts and proteins fasta files. I'm sure I have a configuration setting incorrect or one of the dependencies not correctly installed, but I can't figure out what the problem is. Any thoughts on how I can resolve this issue and generate these files? Ideally I would love to be able to generate these files without having to run the whole pipeline again. Details on my configuration settings and the contents of the run.log file from my example above are pasted below.
> 
> Thank you,
> Kevin
> 
> -----
> run.log from the example folder above looks like this:
> -----
> SHARED_ID    d574e9ca9b0019a9fe147ccb9db3588b
> CTL_OPTIONS    maker_gff
> CTL_OPTIONS    other_gff
> CTL_OPTIONS    est    test-transcriptome.fa
> CTL_OPTIONS    est_reads
> CTL_OPTIONS    altest    KK273.fa
> CTL_OPTIONS    est_gff
> CTL_OPTIONS    altest_gff
> CTL_OPTIONS    protein    test-AA.fa
> CTL_OPTIONS    protein_gff
> CTL_OPTIONS    model_org    all
> CTL_OPTIONS    repeat_protein    te_proteins.fasta
> CTL_OPTIONS    rmlib
> CTL_OPTIONS    rm_gff
> CTL_OPTIONS    organism_type    eukaryotic
> CTL_OPTIONS    predictor    est2genome,genemark,protein2genome
> CTL_OPTIONS    est2genome    1
> CTL_OPTIONS    altest2genome    0
> CTL_OPTIONS    snaphmm
> CTL_OPTIONS    gmhmm    output/gmhmm.mod
> CTL_OPTIONS    augustus_species
> CTL_OPTIONS    fgenesh_par_file
> CTL_OPTIONS    model_gff
> CTL_OPTIONS    pred_gff
> CTL_OPTIONS    max_dna_len    100000
> CTL_OPTIONS    split_hit    10000
> CTL_OPTIONS    pred_flank    200
> CTL_OPTIONS    pred_stats    0
> CTL_OPTIONS    min_protein    0
> CTL_OPTIONS    AED_threshold    1
> CTL_OPTIONS    single_exon    0
> CTL_OPTIONS    single_length    250
> CTL_OPTIONS    keep_preds    0
> CTL_OPTIONS    map_forward    0
> CTL_OPTIONS    est_forward    0
> CTL_OPTIONS    correct_est_fusion    0
> CTL_OPTIONS    alt_splice    0
> CTL_OPTIONS    always_complete    0
> CTL_OPTIONS    alt_peptide    C
> CTL_OPTIONS    evaluate    0
> CTL_OPTIONS    blast_type    ncbi+
> CTL_OPTIONS    softmask    1
> CTL_OPTIONS    pcov_blastn    0.8
> CTL_OPTIONS    pid_blastn    0.85
> CTL_OPTIONS    eval_blastn    1e-10
> CTL_OPTIONS    bit_blastn    40
> CTL_OPTIONS    depth_blastn    0
> CTL_OPTIONS    pcov_rm_blastx    0.5
> CTL_OPTIONS    pid_rm_blastx    0.4
> CTL_OPTIONS    eval_rm_blastx    1e-06
> CTL_OPTIONS    bit_rm_blastx    30
> CTL_OPTIONS    pcov_blastx    0.5
> CTL_OPTIONS    pid_blastx    0.4
> CTL_OPTIONS    depth_blastx    0
> CTL_OPTIONS    eval_blastx    1e-06
> CTL_OPTIONS    bit_blastx    30
> CTL_OPTIONS    pcov_tblastx    0.8
> CTL_OPTIONS    pid_tblastx    0.85
> CTL_OPTIONS    eval_tblastx    1e-10
> CTL_OPTIONS    bit_tblastx    40
> CTL_OPTIONS    depth_tblastx    0
> CTL_OPTIONS    ep_score_limit    20
> CTL_OPTIONS    en_score_limit    20
> CTL_OPTIONS    enable_fathom    0
> CTL_OPTIONS    unmask    0
> CTL_OPTIONS    model_pass    0
> CTL_OPTIONS    est_pass    0
> CTL_OPTIONS    altest_pass    0
> CTL_OPTIONS    protein_pass    0
> CTL_OPTIONS    rm_pass    0
> CTL_OPTIONS    other_pass    0
> CTL_OPTIONS    pred_pass    0
> CTL_OPTIONS    run    genemark
> LOGCHILD /media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/run.log.child.0 
> LOGCHILD /media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/run.log.child.0 
> LOGCHILD /media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/run.log.child.0 
> STARTED test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/scaffold-334630.abinit_nomask.0.gmhmm%2Emod.genemark 
> FINISHED test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/scaffold-334630.abinit_nomask.0.gmhmm%2Emod.genemark 
> STARTED test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/scaffold-334630.0.pred.raw.section 
> FINISHED test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/scaffold-334630.0.pred.raw.section 
> LOGCHILD /media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/run.log.child.0 
> LOGCHILD /media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/run.log.child.0 
> LOGCHILD /media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/run.log.child.0 
> STARTED test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/scaffold-334630.0.final.section 
> FINISHED test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/scaffold-334630.0.final.section 
> LOGCHILD /media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/run.log.child.0 
> LOGCHILD /media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/run.log.child.0 
> LOGCHILD /media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.maker.output/test_scaffolds_annotated_as_metazoan_by_MG-RAST_datastore/00/0C/scaffold-334630//theVoid.scaffold-334630/run.log.child.0 
> 
> -----
> maker_opts
> -----
> #-----Genome (these are always required)
> genome=/media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test_scaffolds_annotated_as_metazoan_by_MG-RAST.fas #genome sequence (fasta file or fasta embeded in GFF3 file)
> organism_type=eukaryotic #eukaryotic or prokaryotic. Default is eukaryotic
> 
> #-----Re-annotation Using MAKER Derived GFF3
> maker_gff= #MAKER derived GFF3 file
> est_pass=0 #use ESTs in maker_gff: 1 = yes, 0 = no
> altest_pass=0 #use alternate organism ESTs in maker_gff: 1 = yes, 0 = no
> protein_pass=0 #use protein alignments in maker_gff: 1 = yes, 0 = no
> rm_pass=0 #use repeats in maker_gff: 1 = yes, 0 = no
> model_pass=0 #use gene models in maker_gff: 1 = yes, 0 = no
> pred_pass=0 #use ab-initio predictions in maker_gff: 1 = yes, 0 = no
> other_pass=0 #passthrough anyything else in maker_gff: 1 = yes, 0 = no
> 
> #-----EST Evidence (for best results provide a file for at least one)
> est=/media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test-transcriptome.fa #set of ESTs or assembled mRNA-seq in fasta format
> altest=/media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/KK273.fa #EST/cDNA sequence file in fasta format from an alternate organism
> est_gff= #aligned ESTs or mRNA-seq from an external GFF3 file
> altest_gff= #aligned ESTs from a closly relate species in GFF3 format
> 
> #-----Protein Homology Evidence (for best results provide a file for at least one)
> protein=/media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/test-AA.fa #protein sequence file in fasta format (i.e. from mutiple oransisms)
> protein_gff=  #aligned protein homology evidence from an external GFF3 file
> 
> #-----Repeat Masking (leave values blank to skip repeat masking)
> model_org=all #select a model organism for RepBase masking in RepeatMasker
> rmlib= #provide an organism specific repeat library in fasta format for RepeatMasker
> repeat_protein=/usr/local/bin/maker/data/te_proteins.fasta #provide a fasta file of transposable element proteins for RepeatRunner
> rm_gff= #pre-identified repeat elements from an external GFF3 file
> prok_rm=0 #forces MAKER to repeatmask prokaryotes (no reason to change this), 1 = yes, 0 = no
> softmask=1 #use soft-masking rather than hard-masking in BLAST (i.e. seg and dust filtering)
> 
> #-----Gene Prediction
> snaphmm= #SNAP HMM file
> gmhmm=/media/kmkocot/Sclerite/genome_projects/test/Ray_2_assembly/MAKER/output/gmhmm.mod #GeneMark HMM file
> augustus_species= #Augustus gene prediction species model
> fgenesh_par_file= #FGENESH parameter file
> pred_gff= #ab-initio predictions from an external GFF3 file
> model_gff= #annotated gene models from an external GFF3 file (annotation pass-through)
> est2genome=1 #infer gene predictions directly from ESTs, 1 = yes, 0 = no
> protein2genome=1 #infer predictions from protein homology, 1 = yes, 0 = no
> trna=0 #find tRNAs with tRNAscan, 1 = yes, 0 = no
> snoscan_rrna= #rRNA file to have Snoscan find snoRNAs
> unmask=0 #also run ab-initio prediction programs on unmasked sequence, 1 = yes, 0 = no
> 
> #-----Other Annotation Feature Types (features MAKER doesn't recognize)
> other_gff= #extra features to pass-through to final MAKER generated GFF3 file
> 
> #-----External Application Behavior Options
> alt_peptide=C #amino acid used to replace non-standard amino acids in BLAST databases
> cpus=1 #max number of cpus to use in BLAST and RepeatMasker (not for MPI, leave 1 when using MPI)
> 
> #-----MAKER Behavior Options
> max_dna_len=100000 #length for dividing up contigs into chunks (increases/decreases memory usage)
> min_contig=1 #skip genome contigs below this length (under 10kb are often useless)
> 
> pred_flank=200 #flank for extending evidence clusters sent to gene predictors
> pred_stats=0 #report AED and QI statistics for all predictions as well as models
> AED_threshold=1 #Maximum Annotation Edit Distance allowed (bound by 0 and 1)
> min_protein=0 #require at least this many amino acids in predicted proteins
> alt_splice=0 #Take extra steps to try and find alternative splicing, 1 = yes, 0 = no
> always_complete=0 #extra steps to force start and stop codons, 1 = yes, 0 = no
> map_forward=0 #map names and attributes forward from old GFF3 genes, 1 = yes, 0 = no
> keep_preds=0 #Concordance threshold to add unsupported gene prediction (bound by 0 and 1)
> 
> split_hit=10000 #length for the splitting of hits (expected max intron size for evidence alignments)
> single_exon=0 #consider single exon EST evidence when generating annotations, 1 = yes, 0 = no
> single_length=250 #min length required for single exon ESTs if 'single_exon is enabled'
> correct_est_fusion=0 #limits use of ESTs in annotation to avoid fusion genes
> 
> tries=2 #number of times to try a contig if there is a failure for some reason
> clean_try=0 #remove all data from previous run before retrying, 1 = yes, 0 = no
> clean_up=0 #removes theVoid directory with individual analysis files, 1 = yes, 0 = no
> TMP= #specify a directory other than the system default temporary directory for temporary files
> 
> -- 
> Kevin M. Kocot, Ph.D.
> NSF International Postdoctoral Research Fellow
> Degnan Lab
> The University of Queensland
> School of Biological Sciences
> 325 Goddard Building 8
> St. Lucia, QLD 4072
> Australia
> Ph: +61 0402 488 430
> 
> 
> _______________________________________________
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