We are having the exact same issue, on the main server and our (recent)
cloud instances.

Were some of the hidden Cuffdiff parameters modified since fall 2012?

Cheers,
Mo Heydarian
On Mar 13, 2013 11:02 AM, "Jenna Smith" <jes...@case.edu> wrote:

> Hi,
>
> I'll preface my concern by saying that I'm a novice to Cufflinks.  Back in
> September, I performed a Cuffdiff analysis comparing a wild-type and mutant
> condition.  The analysis returned ~800 transcripts differentially regulated
> between the two with statistical significance.  Recently, I've rerun the
> Cuffdiff analysis - using exactly the same files stored in Galaxy for all
> inputs, and with all the same parameters - and only get a few dozen
> statistically significant hits.  However, all of the data besides the p and
> q values are essentially identical between these two runs, so I am really
> unclear as to what is causing the difference.  Here is just one clear
> example:
>
> From run 1:
> YFR026C
> FPKM 1 = 17.2434
> FPKM 2 = 196.735
> log2(fold change) = 3.51214
> p = 1.64E-8
> q = 7.33E-6
> significant = yes
>
> From run 2:
> YFR026C
> FPKM 1 = 14.4489
> FPKM 2 = 144.939
> log2(fold change) = 3.32641
> p = 0.000170034
> q = 0.0719964
> significant = no
>
> The second Cuffdiff analysis shows there is still a ~10-fold difference
> between conditions, but this is not statistically significant.  Has the
> version of Cuffdiff on Galaxy been updated such that some parameters have
> changed, that could explain this difference?  Or, is there some setting I
> am missing that would cause very large changes to fail statistical
> significance testing?  Any help or input would be appreciated, I am really
> at a loss for why executing what should be exactly the same task is giving
> vastly different results.  I could just be overlooking something very
> fundamental that is obvious to someone with more experience with this
> program.  Thanks.
>
> -Jenna Smith
>
>
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