Re: [galaxy-user] RNA-seq analysis withtout reference genome

2012-10-08 Thread Jennifer Jackson
Hello Alicia, There are two tools in the main Galaxy Tool Shed (http://toolshed.g2.bx.psu.edu/) that would likely be helpful. Read the input requirements for each to decide which is a better fit. Search for 'deseq' to find them: deseq_and_sam2counts deseq_hts To use these tools, a local

[galaxy-user] RNA-seq analysis withtout reference genome

2012-10-05 Thread Alicia R . Pérez-Porro
Hi all, I'm about to start doing gene expression analysis but i don't have a reference genome. I got my sequences from Illumina and i did my assemblies with CLC bio. I was planning to use DESeq for the analysis but for that i need my gene expression count values. There is any tool in Galaxy that i

[galaxy-user] RNA-Seq analysis

2012-02-06 Thread 杨继文
Since the pictures are too big for the mailling list. I will upload it in seperate emails. Dear all, I am using Galaxy for RNA-Seq analysis. I expect two lists: differentially expressed transcripts and differentially expressed genes. In these two lists, I would like to see the gene name, gene

Re: [galaxy-user] RNA seq analysis

2011-05-07 Thread puvan001
Hi Thank you! yes, your guess is correct. Now it works. Sumathy On May 7 2011, Jeremy Goecks wrote: Sumathy, It sounds like you're on the right track. To visualize data for a custom build in Trackster, you need to create a custom build and use that in Trackster: (1) using the top

Re: [galaxy-user] RNA seq analysis

2011-05-07 Thread Jeremy Goecks
Sumathy, It sounds like you're on the right track. To visualize data for a custom build in Trackster, you need to create a custom build and use that in Trackster: (1) using the top tabs in Galaxy, go to User --> Custom Builds; (2) add a new build with the length info as follows: Important not

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread vasu punj
Thanks Jim,   Vasu   --- On Fri, 5/6/11, Jim Robinson wrote: From: Jim Robinson Subject: Re: [galaxy-user] RNA seq analysis To: "vasu punj" Cc: "Austin Paul" , "Sean Davis" , "galaxy-user@lists.bx.psu.edu" , "puvan...@umn.edu" Date: Friday,

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread puvan001
great work but How hard is to include index functionality with in IGV. I know we can use sam tools also but just for convinence if it is not that much of work. Vasu --- On Fri, 5/6/11, Sean Davis wrote: From: Sean Davis Subject: Re: [galaxy-user] RNA seq analysis To: "Austin Paul&quo

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread puvan001
Hi I may be doing in a wrong way. I clicked trackster and I added the custom build genome. Since it is a very small genome (~2kb), I considered this as a single contig. Then I cliked "add tracks" and added my data file. But I got a message "no data for this contig. Whenever I used built in g

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread Jim Robinson
have been using IGV 2 beta version it is great work but How hard is to include index functionality with in IGV. I know we can use sam tools also but just for convinence if it is not that much of work. Vasu --- On Fri, 5/6/11, Sean Davis wrote: From: Sean Davis Subject: Re: [galaxy-use

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread Jeremy Goecks
Sumathy, What kind of problems are you having with Trackster? J. On May 6, 2011, at 8:30 PM, wrote: > Hello > > I was able to run RNA seq data against a custom build genome. How can I > visualize the results. I tried via trackster and unfortunately I couldn't. > Can you help me? > > > Tha

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread vasu punj
convinence if it is not that much of work. Vasu --- On Fri, 5/6/11, Sean Davis wrote: From: Sean Davis Subject: Re: [galaxy-user] RNA seq analysis To: "Austin Paul" Cc: "galaxy-user@lists.bx.psu.edu" , "puvan...@umn.edu" Date: Friday, May 6, 2011, 8:02 PM IGV

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread vasu punj
I generally take the GTF file to UCSC genome browser. If you are visualizing Bam file after alignment. I found IGV convinenet, though you may be able to visualize in Galaxy.   Vasu --- On Fri, 5/6/11, puvan...@umn.edu wrote: From: puvan...@umn.edu Subject: Re: [galaxy-user] RNA seq analysis

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread Austin Paul
Oops. Good to know. Thanks. Austin On Fri, May 6, 2011 at 6:02 PM, Sean Davis wrote: > IGV reads BAM files just fine; no need to convert to SAM. > > Sean > > On Fri, May 6, 2011 at 8:45 PM, Austin Paul wrote: > >> There are many ways. I typically use IGV. It needs a sam file, so I >> first

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread Sean Davis
IGV reads BAM files just fine; no need to convert to SAM. Sean On Fri, May 6, 2011 at 8:45 PM, Austin Paul wrote: > There are many ways. I typically use IGV. It needs a sam file, so I first > convert the bam to sam in galaxy, then download the sam file. In IGV, I > upload the reference and t

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread Austin Paul
There are many ways. I typically use IGV. It needs a sam file, so I first convert the bam to sam in galaxy, then download the sam file. In IGV, I upload the reference and the sam file, then use IGVtools to index the sam file, then I can visualize the data. Austin On Fri, May 6, 2011 at 5:30 PM,

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread puvan001
Hello I was able to run RNA seq data against a custom build genome. How can I visualize the results. I tried via trackster and unfortunately I couldn't. Can you help me? Thanks Sumathy ___ The Galaxy User list should be used for the di

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread puvan001
Hi Austin I did all these (grooming and trimming)on rna-seq data and I don't have a problem with built in genome . I'll try again! Thanks Sumathy On May 6 2011, Austin Paul wrote: Hi, You need to run fastq groomer on your rna-seq data. Your reference is fine as a fasta. Austin On

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread Austin Paul
Hi, You need to run fastq groomer on your rna-seq data. Your reference is fine as a fasta. Austin On Fri, May 6, 2011 at 10:26 AM, wrote: > > Hi David, > > Thanks!When I tried to run Tophat, it doesn't recognise my FASTA file and > it says "History does not include a dataset of the required f

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread puvan001
Hi David, Thanks!When I tried to run Tophat, it doesn't recognise my FASTA file and it says "History does not include a dataset of the required format / build". Do you have any thoughts about this? Now it makes more sense about "multihits". Thanks for sharing your workflow. With regards

Re: [galaxy-user] RNA seq analysis

2011-05-06 Thread David Matthews
Hi, I have done exactly the same kind of thing for adenovirus so I can help with it. In answer to question 1 you do not need to index it will be done for you when tophat is called. Secondly you should leave the 40 multihits as it is and post analysis filter out the multihits - this will allow y

[galaxy-user] RNA seq analysis

2011-05-06 Thread puvan001
Hi I have a couple of questions regarding RNA seq analysis. My questions are 1.I need to use a viral genome (very small, ~2kb ) as a reference genome and it is not available in Galaxy. I guess I can use this data from my history. I have a fasta file but I am not sure whether I have to do some

Re: [galaxy-user] RNA seq analysis

2011-05-03 Thread Jennifer Jackson
Hello, On 4/28/11 9:21 PM, puvan...@umn.edu wrote: I am new to Galaxy and I am not sure whether these topics were discussed earlier. I followed the steps up to cufflinks and I did not have any problems. Thanks for the RNA seq tutorial. My questions are 1. How do I know the number of reads mapped

[galaxy-user] RNA seq analysis

2011-04-28 Thread puvan001
I am new to Galaxy and I am not sure whether these topics were discussed earlier. I followed the steps up to cufflinks and I did not have any problems. Thanks for the RNA seq tutorial. My questions are 1. How do I know the number of reads mapped against the reference genome used after Top Hat ma

Re: [galaxy-user] RNA seq analysis and GTF files

2011-04-08 Thread David K Crossman
ail address. Thanks, David From: Jeremy Goecks [mailto:jeremy.goe...@emory.edu] Sent: Thursday, April 07, 2011 3:42 PM To: David K Crossman Cc: galaxy-user Subject: Re: [galaxy-user] RNA seq analysis and GTF files David, can you please share your history with me and I'll take a look (H

Re: [galaxy-user] RNA seq analysis and GTF files

2011-04-07 Thread Jeremy Goecks
t before moving on). Any info, suggestions or help would be greatly appreciated. Thanks, David -Original Message- From: galaxy-user-boun...@lists.bx.psu.edu [mailto:galaxy-user-boun...@lists.bx.psu.edu ] On Behalf Of Jeremy Goecks Sent: Friday, April 01, 2011 8:47 AM To: Cc: gala

Re: [galaxy-user] RNA seq analysis and GTF files

2011-04-07 Thread David K Crossman
[mailto:galaxy-user-boun...@lists.bx.psu.edu] On Behalf Of Jeremy Goecks Sent: Friday, April 01, 2011 8:47 AM To: Cc: galaxy-user Subject: Re: [galaxy-user] RNA seq analysis and GTF files On Mar 31, 2011, at 12:30 PM, mailto:ssa...@ccib.mgh.harvard.edu>> mailto:ssa...@ccib.mgh.

Re: [galaxy-user] RNA seq analysis and GTF files

2011-04-01 Thread Jeremy Goecks
On Mar 31, 2011, at 12:30 PM, wrote: > Hi Jeremy, > I used your exercise to perform an RNA-seq analysis. First I encountered a > problem where the gene IDs were missing from the results. Jen from the Galaxy > team suggested this: > > "Yes, the team has taken a look and there are a few t

Re: [galaxy-user] RNA seq analysis

2011-02-24 Thread David Matthews
HI Jeremy, Thanks for the feedback. I know what you mean about tophat not having the same functionality of bowtie. However, I think whatever tophat does do (now or in the future) I think it is useful to collect the multihits separately since either you leave them in and over estimate gene expre

Re: [galaxy-user] RNA seq analysis

2011-02-24 Thread Ann Loraine
Hello, I like your approach of running the alignment tools with liberal settings and then filtering the results into different categories. This discussion reminds me of how in expression microarray analysis, we face uncertainty as to what molecules (exactly) are hybridizing to the probes on a chi

Re: [galaxy-user] RNA seq analysis

2011-02-24 Thread David Matthews
Thanks Ann for your comments and for the stuff you showed at IGB - looks very interesting. I agree that multihits may the equivalent of the problem you describe from microarrays. I think, for me anyway, knowing the scale if the issue is the key thing at this stage. As you imply from your email t

Re: [galaxy-user] RNA seq analysis

2011-02-24 Thread Ann Loraine
Hello, I like your approach of running the alignment tools with liberal settings and then filtering the results into different categories. This discussion reminds me of how in expression microarray analysis, we face uncertainty as to what molecules (exactly) are hybridizing to the probes on a ch

Re: [galaxy-user] RNA seq analysis

2011-02-23 Thread vasu punj
  --- On Wed, 2/23/11, Jeremy Goecks wrote: From: Jeremy Goecks Subject: Re: [galaxy-user] RNA seq analysis To: "David Matthews" Cc: galaxy-u...@bx.psu.edu Date: Wednesday, February 23, 2011, 10:05 PM Hi David, This is a really interesting workflow. My comments: (1) I encourage you

Re: [galaxy-user] RNA seq analysis

2011-02-23 Thread Jeremy Goecks
Hi David, This is a really interesting workflow. My comments: (1) I encourage you to start a discussion about this idea on seqanswers.com; you'll reach more people and may have a better discussion there. Ideally, you'll get a Tophat developer to chime in on what I perceive to be the main issue

Re: [galaxy-user] RNA seq analysis

2011-02-23 Thread David Matthews
Hi all, Further to my last email, I've published a workflow (Bristol workflow ) which does what I described below - hope this helps in understanding what I'm on about (!). Best Wishes, David. On 23 Feb 2011, at 14:41, David Matthews wrote: > Hi Jeremy, > > I thought I'd write to get a

[galaxy-user] RNA seq analysis

2011-02-23 Thread David Matthews
Hi Jeremy, I thought I'd write to get a discussion of a workflow for people doing RNA seq that I have found very useful and addresses some issues in mapping mRNA derived RNA-seq paired end data to the genome using tophat. Here is the approach I use (I have a human mRNA sample deep sequenced wit