Question: Hi all,

I am trying to get the cegma working on the sample dataset that came along with the package. I have already installed all the pre-requistes (`ncbi-blast 2.2.25+`, `hmmer 3.0`, `geneid 1.4.4` and `genewise 2.4`). When executed (as suggested in the `README` file), I get the error saying that `geneid-train did not work properly`. Please see the entire `STDOUT` (verbose) below. Any suggestions will be greatly appreciated!

$ cegma -v -ext --genome sample.dna --protein sample.prot -o sample &>sample_data.stdout
$ cat sample_data.stdout

********************************************************************************
** MAPPING PROTEINS TO GENOME (TBLASTN) **
********************************************************************************
RUNNING: genome_map -n genome -p 6 -o 5000 -c 2000 -t 1 -v sample.prot sample.dna 2>sample.cegma.errors

Building a new DB, current time: 05/01/2014 09:10:46
New DB name: /tmp/genome25484.blastdb
New DB title: sample.dna
Sequence type: Nucleotide
Keep Linkouts: T
Keep MBits: T
Maximum file size: 1073741824B
Adding sequences from FASTA; added 1 sequences in 0.07232 seconds.
Processing KOG: KOG0292
Processing KOG: KOG0328
Processing KOG: KOG0466
Processing KOG: KOG0631
Processing KOG: KOG0659
Processing KOG: KOG1458
Processing KOG: KOG1742
Processing KOG: KOG1762
Processing KOG: KOG1769
Processing KOG: KOG1780
Processing KOG: KOG1992
Processing KOG: KOG2067
Processing KOG: KOG2803
Processing KOG: KOG2916
Processing KOG: KOG3180
Processing KOG: KOG3232
Processing KOG: KOG3418
Processing KOG: KOG3497
Found 86 candidate regions in sample.dna

********************************************************************************
** MAKING INITIAL GENE PREDICTIONS FOR CORE GENES (GENEWISE + GENEID) **
********************************************************************************

RUNNING: local_map -n local -f -h /data004/software/GIF/packages/cegma/2.4.010312/data/hmm_profiles -i KOG -v genome.chunks.fa 2>sample.cegma.errors
Processing chunk: KOG0292.1
Processing chunk: KOG0292.6
Processing chunk: KOG0292.7
Processing chunk: KOG0292.2
Processing chunk: KOG0292.4
Processing chunk: KOG0328.7
Processing chunk: KOG0328.18
Processing chunk: KOG0328.16
Processing chunk: KOG0328.15
Processing chunk: KOG0328.8
Processing chunk: KOG0466.5
Processing chunk: KOG0466.4
Processing chunk: KOG0466.3
Processing chunk: KOG0466.2
Processing chunk: KOG0631.10
Processing chunk: KOG0631.6
Processing chunk: KOG0631.9
Processing chunk: KOG0631.5
Processing chunk: KOG0631.12
Processing chunk: KOG0659.19
Processing chunk: KOG0659.7
Processing chunk: KOG0659.20
Processing chunk: KOG0659.21
Processing chunk: KOG0659.10
Processing chunk: KOG1458.6
Processing chunk: KOG1458.8
Processing chunk: KOG1458.5
Processing chunk: KOG1458.3
Processing chunk: KOG1458.9
Processing chunk: KOG1742.8
Processing chunk: KOG1742.3
Processing chunk: KOG1742.9
Processing chunk: KOG1742.6
Processing chunk: KOG1742.5
Processing chunk: KOG1762.3
Processing chunk: KOG1762.2
Processing chunk: KOG1769.22
Processing chunk: KOG1769.15
Processing chunk: KOG1769.8
Processing chunk: KOG1769.5
Processing chunk: KOG1769.27
Processing chunk: KOG1780.5
Processing chunk: KOG1780.2
Processing chunk: KOG1780.7
Processing chunk: KOG1780.3
Processing chunk: KOG1780.6
Processing chunk: KOG1992.1
Processing chunk: KOG1992.8
Processing chunk: KOG1992.2
Processing chunk: KOG1992.3
Processing chunk: KOG1992.6
Processing chunk: KOG2067.6
Processing chunk: KOG2067.7
Processing chunk: KOG2067.3
Processing chunk: KOG2067.9
Processing chunk: KOG2067.8
Processing chunk: KOG2803.7
Processing chunk: KOG2803.5
Processing chunk: KOG2803.8
Processing chunk: KOG2803.4
Processing chunk: KOG2803.6
Processing chunk: KOG2916.9
Processing chunk: KOG2916.4
Processing chunk: KOG2916.11
Processing chunk: KOG2916.10
Processing chunk: KOG2916.2
Processing chunk: KOG3180.12
Processing chunk: KOG3180.10
Processing chunk: KOG3180.3
Processing chunk: KOG3180.7
Processing chunk: KOG3180.9
Processing chunk: KOG3232.6
Processing chunk: KOG3232.2
Processing chunk: KOG3232.4
Processing chunk: KOG3232.3
Processing chunk: KOG3232.5
Processing chunk: KOG3418.12
Processing chunk: KOG3418.10
Processing chunk: KOG3418.3
Processing chunk: KOG3418.9
Processing chunk: KOG3418.8
Processing chunk: KOG3497.9
Processing chunk: KOG3497.1
Processing chunk: KOG3497.10
Processing chunk: KOG3497.2
Processing chunk: KOG3497.11
NOTE: created 23 geneid predictions

********************************************************************************
** FILTERING INITIAL PROTEINS PRODUCED BY GENEID (HMMER) **
********************************************************************************

RUNNING: hmm_select -i KOG -o local -t 1 -v /data004/software/GIF/packages/cegma/2.4.010312/data/hmm_profiles local.geneid.fa /data004/software/GIF/packages/cegma/2.4.010312/data/profiles_cutoff.tbl 2>sample.cegma.errors
Processing geneid prediction: KOG0292.6
Processing geneid prediction: KOG0292.7
Processing geneid prediction: KOG0328.7
Processing geneid prediction: KOG0328.18
Processing geneid prediction: KOG0328.8
Processing geneid prediction: KOG0466.5
Processing geneid prediction: KOG0631.10
Processing geneid prediction: KOG0659.19
Processing geneid prediction: KOG0659.7
Processing geneid prediction: KOG0659.20
Processing geneid prediction: KOG1742.8
Processing geneid prediction: KOG1762.3
Processing geneid prediction: KOG1769.22
Processing geneid prediction: KOG1992.1
Processing geneid prediction: KOG2067.6
Processing geneid prediction: KOG2067.7
Processing geneid prediction: KOG2803.7
Processing geneid prediction: KOG2803.8
Processing geneid prediction: KOG2916.9
Processing geneid prediction: KOG3180.12
Processing geneid prediction: KOG3232.6
Processing geneid prediction: KOG3418.12
Processing geneid prediction: KOG3497.9
NOTE: Found 15 geneid predictions with scores above threshold value

********************************************************************************
** CALCULATING GENEID PARAMETERS FROM SELECTED GENEID PREDICTIONS **
********************************************************************************

RUNNING: geneid-train -v local.geneid.selected.gff local.geneid.selected.dna geneid_params 2>sample.cegma.errors
DATA COLLECTED: 15 Coding sequences containing 48 introns
Intron model
geneid-train did not work properly

EDIT:

So, when I just run the final step:

$ geneid-train -v local.geneid.selected.gff local.geneid.selected.dna geneid_params

I get:

DATA COLLECTED: 15 Coding sequences containing 48 introns
Intron model
Use of uninitialized value in numeric eq (==) at /data004/software/GIF/packages/cegma/2.4.010312/lib/geneid.pm line 264.
some values in Markov model with zero counts, use pseudocounts at /data004/software/GIF/packages/cegma/2.4.010312/lib/geneid.pm line 270.

Answer: Hi again,

I think I (finally) might have a fix. The problem might result from people with newer versions of Perl in which an old Perl function is deprecated (in certain situations).

To test whether this is the case, you'll need to change one line in each of the geneid.pm and HMMstar.pm files (inside the lib folder of the CEGMA installation). Maybe make copies of the originals as well. In geneid.pm change line 108 and in HMMstar.pm change line 41 as follows:

BEFORE: $code .= "$tab foreach my \$c$i qw(A C G T) {\n";
AFTER: $code .= "$tab foreach my \$c$i (qw(A C G T)) {\n";

And let me know if that works!

Here is some more info about how this function changed in Perl 5.14: http://blogs.perl.org/users/rurban/2010/09/qw-in-list-context-deprecated.html