Thanks Jen,

I have not had good luck with the Visualization feature. Using BAM files, it 
has worked once to produce a single histogram but mostly seems stuck preparing 
data for up to 3 hours with no production. Additionally, once a graph is made 
how can one save the graph as a picture file; pdf, etc?

Best,
Trent

From: Jennifer Jackson [mailto:j...@bx.psu.edu]
Sent: Wednesday, May 30, 2012 3:17 PM
To: Fowler, Trent
Cc: galaxy-u...@bx.psu.edu
Subject: Re: [galaxy-user] Genome Browser Histogram Visualization of Accepted 
Hits

Hi Trent,

Hiram's advice is very good for UCSC display (thanks Hiram!)

You most likely heard of converting BAM->SAM->Interval->BED->BigBED. But, Hiram 
is exactly correct -  there will be a great loss of information (notably the 
full individual sequence data itself, as only the coordinates + variation will 
remain, plus the data will be in BED6, which means no splicing, just global 
coordinates - you won't want this for RNA-seq).

Since you are examining RNA-seq data, I also wanted to remind you of the choice 
to use Trackster for visualization (top Galaxy menu -> "Visualization"). A 
local or custom reference genome can be used, histogram or full read display is 
possible, and deeply sequenced areas can be "unpacked" in stages by using the 
visualization controls (really nice). Other features such as drag and drop 
composite track creation, new datasets from selects within Trackster (to feed 
more analysis), strand coloring, etc. are fairly recent enhancements (see: 
http://wiki.g2.bx.psu.edu/DevNewsBriefs/2012_03_12#Galaxy_Track_Browser_.28GTB.29
 
http://wiki.g2.bx.psu.edu/DevNewsBriefs/2012_05_11#Galaxy_Track_Browser_.28GTB.29).
  More undocumented (so far!) features are added all the time, so good to 
experiment. A new paper is in the works that will cover the details. And there 
will be tutorials, screencasts, etc. later on, but for now during the active 
development cycle, just using it is best and is pretty intuitive. We're 
interested in what you like or wish was added (this is true to everyone reading 
this!). No promises, but is good to get feedback!

If there is a particular UCSC track (or another source mapped to same reference 
genome) that you wish to compare your data against, it could be imported into 
Galaxy and added to your Trackster visualization session. There are no known 
limits on data size/depth - only the general Galaxy dataset file size limit of 
50G  (applies to all datasets) - although I suppose an extreme corner case 
could probably be devised/uncovered. The point is that a routine RNA-seq 
experiment shouldn't present a problem. Special tuning to speed up and handle 
depth has been a big area of recent development activity. If you do run into 
issues with Trackster, feel free to ask questions (I'll probably ask you to 
send a share link to the visualization on the pubic main Galaxy instance and 
bring Jeremy into the discussion if the solution is not clear).

Please let us know if you need more help,

Jen

On 5/29/12 11:57 AM, Fowler, Trent wrote:
Hello,

I am attempting to run accepted hit data from Tophat output into the UCSC 
Genome Web Browser for visualization of sequencing hits in specific genes. 
However, the BAM files yield tiles and are too large to present through the 
browser. Is there a better file format to convert to that would allow better 
visualization such as histograms?

>From word of mouth, I have been told to convert BAMs to BEDs and put BED files 
>through the browser. However, I notice that Galaxy does not have an option for 
>this and the oft used BEDtools appears to involve writing code, which is above 
>my computer abilities.

Any tips or solutions on how to obtain histograms from sequencing data would be 
very welcome.

Thanks

Trent Fowler




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Jennifer Jackson

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___________________________________________________________
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