Dear Galaxy users that also use Megan,
I hope you can help me combining Galaxy with Megan4. I followed the Galaxy metagenome workflow (author aun1) on shotgun DNA 454 sequences of two samples that I would like to compare. The results are two nice trees and tables with the lowest taxonomic ranks for the two samples.
Besides the taxonomy I am also interested in the functions and I would like to use Megan4 (with SEED and KEGG classification) for this purpose. I tried to import a Galaxy file (.tabular) with c1 =high quality segment name and c2-c13 =blasthit results and a Galaxy fasta file with the high quality segment names and sequences. This does not work and I receive the message parsing failed, no reads found.
I am interested to know how you solved this issue and which files in the Galaxy metagenome workflow you use to import into Megan4.
Or maybe there is other software that you use to compare suits of functional genes in your metagenome datasets?
Judith van Bleijswijk
I used SICR to call peaks and have following out put files:
1. test.1removed bed
2. control1 removed .bed
3. test w 200 graph
4. test w200 normalized graph
5. test w200-G600 FDR.05 island.bed
6. test w200-G600 FDR .05 island filtered.bed
7. test w200-G600 FDR .05 island filtered normalized.wig
8. test w200-G600 FDR.05 score island
9. test w200-G600 summary island
Which of these files should be used. I think file 5 and 6 are the ones for
visualization as well for annotating with genomic regions. I have read the
original paper but it is not very clear what these out puts mean. Could
some one please guide me what these files mean and what is useful and rest
are intermediate files.
My second question is authors of SICER has emphasized about teh importance
of choosing gap size and window size. gap size they mention 1-3 -5
depending upon the peak distribution, but I see in Galaxy the default is
600 gap size do we need to change it to 1,2- 5 or I am missing something.
Any advise please but I did my searching on net.
have a question about the cuffdiff output "differential expression testing".
For most of you might sound "naive" but I'm new to this field and I
have very little background in statistic.
So, I have compared a control sample with 2 biological replicates
using cuffdiff. I have now about 4000 genes which were tested.
1. How do I extrapolate the genes which are up- or downregulated from the 4000?
2. Is there a FPKM value above which a gene is up- or downregulated?
3.I used excel and sorted the values from highest to smallest:
assuming that control highest value is 200 and the correspondent
treated values is 2, can I say that that gene is downregulated in the
treated ssamples by a 100 fold chnage?
4. Do I have to use at all the p_values given in the output to
extrapolate the most up- or downregualted genes?
I do not have yet cummerbund and I am not very good with R. And I 'm lost!
I am attempting to run Cufflinks on Galaxy main to analyze my E. coli
RNAseq data. I have mapped my reads using an outside program (Genious) and
uploaded the resulting BAM file. I also have uploaded the E. coli
annotations as a gtf file. However when I attempt to run Cufflinks using
my annotations it just stays on "Job is waiting to run" for hours. If I
click on "Edit attributes", I see an error message "Required metadata
values are missing". Does this mean that my files are somehow incomplete
and cufflinks will never run, or do I just need to wait longer? When
searching around the mailing lists I saw others have had issues with
bacteria due to its circular chromosome, and was wondering if this might
somehow be related. Thanks.
In the menu "User" -> "Saved Datasets", all datasets are listed even if
some of them have been deleted permanently by deleting its history.
So, it's possible to copy an deleted dataset in the current history and
that is confusing for users.
Do you have any solution to drop these datasets in "saved datasets" ?
Thanks in advance,
I created a workflow (
https://main.g2.bx.psu.edu/u/vijay-uky/w/rna-seq-analysis ) and uploaded
the reference genome (30 MB FASTA file) and the reads (45B FASTQ file after
running the FASTQ groomer). Once I ran the workflow on the public server,
it will enqueue the needed output (steps) but the job just wouldn't run. I
tried to run the same on a local instance at my lab, it works fine. Was
curious to know if there is some issue with running tools such as TopHat,
Cufflinks and Cuffcompare.
If you could let me know, would greatly appreciate.
I find an error in the galaxy wiki page. please see "Admin/Datatypes/Adding Datatypes" which URL is http://wiki.g2.bx.psu.edu/Admin/Datatypes/Adding%20Datatypes
<datatype extension="foo" type="galaxy.datatypes.tabular.Foobar" display_in_upload="true"/><sniffer type="galaxy.datatypes.tabular.Foobar"/>
I guess there should be ":" between "tbular" and "Foobar" not "."