how complete is the draft assembly? cegma?

[chrishah]

Hi guys,
I am doing de novo assembly and was wondering about how to assess the completeness of the assembly. I thought cegma is a nice way of getting an idea, but I cannot get it to run properly. Any Cegma users here who could help me with the error below? Do you maybe have other suggestions, or could point me to an older thread dealing with this issue..

This is what cegma says when running the following: cegma -g test.fa


********************************************************************************
** MAPPING PROTEINS TO GENOME (TBLASTN) **
********************************************************************************

RUNNING: genome_map -n genome -p 6 -o 5000 -c 2000 -t 1 ./data/kogs.fa test.fa 2>output.cegma.errors
FATAL ERROR when running genome_map 32512: ""

Much obliged!

cheers,
Christoph

[Ole]

Hi Christoph,
we're also looking into using Cegma as a step in a validation process of an assembly. It was not easy to get running, and I still don't have it up, but I might have some pointers.

Have you looked at the output.cegma.errors file? I usually run into problems with missing programs, in my case it's the lack of a BLAST+ installation (and I couldn't get BLAST+ to find my correct Boost library, so I am not able to install that myself).

Sincerely,
Ole

[chrishah]

Hei Ole,

I managed to get the cegma up and running by now. It was quite a struggle and with considerable help from Keith Bradnam. Also apparently it is much easier with the 2.4 version of cegma.
The attached file shows roughly what I did.

By the way, I also requested the cegma installation on the UIO cluster, so with access to titan you can use it. you just have to load the appropriate modules..
module load hmmer/3.0 ## thats important! default is hmmer 2.3 and cegma needs v 3
module load geneid
module load blast
module load cegma

and off you go...

hope that helps!

cheers,
Christoph

[StuartG1]

Christoph,
thanks very much for the installation guide. Last year I gave up trying to get cegma running, but using your notes had in installed and working in <1hr
thanks again,
Stuart

[StuartG1]

FATAL ERROR when running genome_map 32512: ""

I also had this error, which was solved (at least in my case) by fixing the fasta headers:

>scaffold1|size2472247
changed to
>scaffold1-size2472247

Stuart

[bhoomi_1789]

i am getting this error.. pls help me out

*******************************************************************************
** MAPPING PROTEINS TO GENOME (TBLASTN) **
********************************************************************************

RUNNING: genome_map -n genome -p 6 -o 5000 -c 2000 -t 1 gms.prot vavaria.dna 2>vav.cegma.errors


Building a new DB, current time: 04/25/2012 21:23:36
New DB name: /tmp/genome4881.blastdb
New DB title: vavaria.dna
Sequence type: Nucleotide
Keep Linkouts: T
Keep MBits: T
Maximum file size: 1073741824B
Adding sequences from FASTA; added 61 sequences in 0.0710599 seconds.
FATAL ERROR when running genome_map 65280: ""

[bryand]

You can get a lot more useful information out of cegma if you look in the error log (in your case, "vav.cegma.errors").

[Robson]

Can't download geneid

Hey guys,

I'm also trying to get CEGMA to work, but I run into problems much earlier than you. My problem is that I can't access geneid from geneid's webpage. Seems like a server problem to me. Is there someone who may send me a zip-file with the linux64 version?

[kbradnam]

Just to confirm StuartG1's advice that CEGMA will fail if any FASTA header contains a pipe character |

This is not the problem of CEGMA itself, but the NCBI blast+ command that makes database files from FASTA files.

Keith (co-developer of CEGMA)

[lilicano]

Hi all,

I followed christoph installation log file for cegma 2.4 which is very well explained.

http://korflab.ucdavis.edu/datasets/...ructions_2.txt

I followed your log file and tried to install cegma 2.5 in an ubuntu machine but got the same error of genome_map not found.

Note: my fasta headers are ok (without weird characters such "|")

copied to /etc/perl
- FAlite.pm
- Cegma.pm

installed
- wise
- wise-doc
- wise2.2.3-rc7
- geneid_v1.4.4
- blast+2.2.30
- hmmer3.0

also set the environment for
~./profile
PATH=$PATH:~/geneid/bin/./
export PATH

PATH=$PATH:~/ncbi-blast-2.2.30+/bin/./
export PATH

PATH=$PATH:~/hmmer-3.0/bin/./
export PATH

PATH=$PATH:~/CEGMA_v2.5/bin/./
export PATH

~./bashrc
export CEGMA=/home/wrparks/Applications/CEGMA_v2.5
export PERL5LIB="/usr/lib/perl/5.18:/home/wrparks/Applications/CEGMA_v2.5/lib"
export WISECONFIGDIR=/home/wrparks/Applications/wise2.2.3-rc7/wisecfg

so if a run cegma inside the bin folder as ./cegma it seems to be running ok as shown below but...
wrparks@wrparks-Precision-T7610:~/Applications/CEGMA_v2.5/bin$ ./cegma
cegma


PROGRAM:
cegma - 2.5

Core Eukaryotic Genes Mapping Approach

----
....but when i tried to run my file it fails and gives me the genome_map not found error as shown below:

wrparks@wrparks-Precision-T7610:~/Applications/CEGMA_v2.5/bin$ ./cegma --genome Ph_HDM502AA_DNAassbly_velvetk71_filt500contigs_2905.fan --output Ph_HDM502AA_DNAassbly_velvetk71_filt500contigs_2905_cegma.output


********************************************************************************
** MAPPING PROTEINS TO GENOME (TBLASTN) **
********************************************************************************

RUNNING: genome_map -n genome -p 6 -o 5000 -c 2000 -t 1 ./data/kogs.fa Ph_HDM502AA_DNAassbly_velvetk71_filt500contigs_2905.fan 2>Ph_HDM502AA_DNAassbly_velvetk71_filt500contigs_2905_cegma.output.cegma.errors
FATAL ERROR when running genome_map 512: ""


Ending CEGMA

wrparks@wrparks-Precision-T7610:~/Applications/CEGMA_v2.5/bin$ ls


any suggestions would be greatly appreciated!

Lili

[kbradnam]

Can you run CEGMA with the supplied sample data to check whether the problem is with your CEGMA installation or with your sequence data file?

[whataBamBam]

I think it's some missing dependencies to CEGMA. I got this error recently.

On my cluster you use source command to get some software in your path..

I did

source cegma-2.5
source blast-2.2.30

then ran my CEGMA command and got the error "FATAL ERROR when running genome_map 6400:"

Then I realised I was missing dependencies so I did:

source hmmer-3.1b1
source blast-2.2.29
source geneid-1.4.4
source wise-2.4.1
source cegma-2.5

wise-2.4.1 is GeneWise

Ran my CEGMA command with no error.

So it looks like you might be missing hmmer or geneid, or genewise or some combination of those three.

My command is

cegma --genome mygenome.fa -o cegma_out

[lilicano]

Hi kbradnam,

Thanks for your suggestion. Yes I run the sample/sample.dna and sample/sample.prot without any error.

Thanks also to whataBamBam for the suggestion, with testing the sample files i noticed the sources must be ok.

Then I got another suggestion from a colleague that maybe cegma was not finding my files so what I did was to create a folder within cegma where i placed my fasta genome file like

“/Applications/CEGMAv2.5/assemblies” where i put my assembly fasta file and run it from /Applications/CEGMAv2.5/bin and then it was ok.

Thanks,

Lili