Hello, Problem: Cuffmerge loses p_id from reference genome in merged gtf file on Galaxy online server resulting in blank cds cuffdiff files. DATA Input to Cuffmerge or Cuffcompare: Two fly cufflinks transcript assemblies created from bam files in Galaxy server using reference annotation and bias correction. Options: Use Reference Annotation: YES UCSC DM3 genes gtf (D. melanogaster) or ENSMBL 5.25 genes gtf Use Sequence Data: YES Result: tss_id found in cuffmerge but no p_id with either reference annotation file. Examples included below. Reference: 2L protein_coding stop_codon 8608 8610 . + 0 exon_number "2"; gene_id "FBgn0031208"; gene_name "CG11023"; p_id "P13746"; transcript_id "FBtr0300689"; transcript_name "CG11023-RB"; tss_id "TSS8369"; Cuffmerge: Cufflinks exon 8193 9484 . + . gene_id "XLOC_000001"; transcript_id "TCONS_00000001"; exon_number "2"; gene_name "CG11023"; oId "FBtr0300689"; nearest_ref "FBtr0300689"; class_code "="; tss_id "TSS1"; 2L Cufflinks exon 66721 67003 . + . gene_id "XLOC_000002"; transcript_id "TCONS_00000003"; exon_number "1"; gene_name "dbr"; oId "CUFF.1.1"; nearest_ref "FBtr0078100"; class_code "j"; tss_id "TSS2"; Help is much appreciated, Thanks!
Hello Christopher, I see that this was also posted at SeqAnswers yesterday. I searched and didn't see anything specifically in the manual or at seqanswers that addresses this (but if someone else can find it, please add to this thread): http://seqanswers.com/forums/showthread.php?t=20084 Do you think this is a problem with the Cuffmerge program itself? If so, going to the authors is the next step by emailing tophat.cufflinks@gmail.com http://wiki.g2.bx.psu.edu/Support#Interpreting_scientific_results (See: /Example/: *unexpected results with* /*RNA-seq analysis*/ *tools.)* * *If you think this is a problem may be with the Galaxy wrapper around CuffMerge, we can take a closer look. Please share a link to your history containing this data. Use "Options (top 'gear' icon in history panel) -> Share or publish", generate a share link, copy link, and email link back directly to me. Note the dataset #'s involved and leave all datasets unhidden/undeleted. Thanks! Jen Galaxy team On 5/15/12 1:38 PM, Christopher M. Weber wrote:
Hello,
Problem: Cuffmerge loses p_id from reference genome in merged gtf file on Galaxy online server resulting in blank cds cuffdiff files.
DATA Input to Cuffmerge or Cuffcompare: Two fly cufflinks transcript assemblies created from bam files in Galaxy server using reference annotation and bias correction.
Options: Use Reference Annotation: YES UCSC DM3 genes gtf (D. melanogaster) or ENSMBL 5.25 genes gtf
Use Sequence Data: YES
Result: tss_id found in cuffmerge but no p_id with either reference annotation file. Examples included below.
Reference: 2L protein_coding stop_codon 8608 8610 . + 0 exon_number "2"; gene_id "FBgn0031208"; gene_name "CG11023"; p_id "P13746"; transcript_id "FBtr0300689"; transcript_name "CG11023-RB"; tss_id "TSS8369";
Cuffmerge: Cufflinks exon 8193 9484 . + . gene_id "XLOC_000001"; transcript_id "TCONS_00000001"; exon_number "2"; gene_name "CG11023"; oId "FBtr0300689"; nearest_ref "FBtr0300689"; class_code "="; tss_id "TSS1"; 2L Cufflinks exon 66721 67003 . + . gene_id "XLOC_000002"; transcript_id "TCONS_00000003"; exon_number "1"; gene_name "dbr"; oId "CUFF.1.1"; nearest_ref "FBtr0078100"; class_code "j"; tss_id "TSS2";
Help is much appreciated,
Thanks!
___________________________________________________________ The Galaxy User list should be used for the discussion of Galaxy analysis and other features on the public server at usegalaxy.org. Please keep all replies on the list by using "reply all" in your mail client. For discussion of local Galaxy instances and the Galaxy source code, please use the Galaxy Development list:
http://lists.bx.psu.edu/listinfo/galaxy-dev
To manage your subscriptions to this and other Galaxy lists, please use the interface at:
-- Jennifer Jackson http://galaxyproject.org
Hello Christopher, We did some testing here using your available data and are fairly certain that the problem is with the tool itself and not the Galaxy wrapper. Contacting the author and asking about the issue is probably the best way forward. If you have time, we would be glad to learn about the outcome. Best, Jen Galaxy team On 5/15/12 1:38 PM, Christopher M. Weber wrote:
Hello,
Problem: Cuffmerge loses p_id from reference genome in merged gtf file on Galaxy online server resulting in blank cds cuffdiff files.
DATA Input to Cuffmerge or Cuffcompare: Two fly cufflinks transcript assemblies created from bam files in Galaxy server using reference annotation and bias correction.
Options: Use Reference Annotation: YES UCSC DM3 genes gtf (D. melanogaster) or ENSMBL 5.25 genes gtf
Use Sequence Data: YES
Result: tss_id found in cuffmerge but no p_id with either reference annotation file. Examples included below.
Reference: 2L protein_coding stop_codon 8608 8610 . + 0 exon_number "2"; gene_id "FBgn0031208"; gene_name "CG11023"; p_id "P13746"; transcript_id "FBtr0300689"; transcript_name "CG11023-RB"; tss_id "TSS8369";
Cuffmerge: Cufflinks exon 8193 9484 . + . gene_id "XLOC_000001"; transcript_id "TCONS_00000001"; exon_number "2"; gene_name "CG11023"; oId "FBtr0300689"; nearest_ref "FBtr0300689"; class_code "="; tss_id "TSS1"; 2L Cufflinks exon 66721 67003 . + . gene_id "XLOC_000002"; transcript_id "TCONS_00000003"; exon_number "1"; gene_name "dbr"; oId "CUFF.1.1"; nearest_ref "FBtr0078100"; class_code "j"; tss_id "TSS2";
Help is much appreciated,
Thanks!
___________________________________________________________ The Galaxy User list should be used for the discussion of Galaxy analysis and other features on the public server at usegalaxy.org. Please keep all replies on the list by using "reply all" in your mail client. For discussion of local Galaxy instances and the Galaxy source code, please use the Galaxy Development list:
http://lists.bx.psu.edu/listinfo/galaxy-dev
To manage your subscriptions to this and other Galaxy lists, please use the interface at:
-- Jennifer Jackson http://galaxyproject.org
participants (2)
-
Christopher M. Weber -
Jennifer Jackson