Hi Allen (we should keep this conversation on the list).
yes, normalized intensity 1 and 2 would be a good place to start for
making MA plots.
If you haven't made MA plots before, i'd suggest looking through the
limma user guide.
I'd recommend making a pdf file with 3x3 rows and columns, then
plotting all 83 of them (thus 10 pages of output).
In addition, a boxplot of the ratios (in log2 space) will reveal
whether these ratios have similar distributions.
cheers,
Mark
On 16/06/2008, at 10:18 AM, ss wrote:
> Hi Mark,
>
> Thanks much! So I should get started with normalized intensity 1 and
> intensity2, right?
> There are 83 arrays and what is the best way to assess MA plot? One
> by one?
>
> Best,
> Allen
>
> On Sun, Jun 15, 2008 at 7:41 PM, Mark Cowley <m.cowley0@gmail.com>
> wrote:
> Hi Allen,
>
> Since you also have intensity 1 and intensity 2 columns, you can
> perform MA plots of your data; for me, this would be the most
> revealing way of identifying how normalised the data is.
>
> Boxplots, or density plots of the ratios should give you a good idea
> of whether the ratios are symmetrically distributed
>
> cheers,
> Mark
>
>
> On 14/06/2008, at 3:32 PM, ss wrote:
>
> Kasper,
>
> Thanks!
>
> I checked and found that:
>
> Ratio = Intensity2/Intensity1
>
> Log(Ratio) is based on 10
>
> Since the data is already being processed, the quick and simple way
to
> verify it is has been
> normalized could be to take the average of each array and see if
> they are
> very similar?
>
> I appreciate!
>
> Allen
>
> On Sat, Jun 14, 2008 at 1:08 AM, Kasper Daniel Hansen <
> khansen@stat.berkeley.edu> wrote:
>
> As you said yourself, these are processed intensities, so how should
> we
> know what to do with them. It does not come out of Agilents
software.
> But given these columns names I guess that
> - Ratio is equal to Intensity 1 / Intensity 2 (or vice verse)
> - Log(Ratio) is the logarithm of Ratio, check yourself to see what
> base
> they are using. My guess is either 2 or 10 but it could be e
> - I have no idea what Fold Change is, my guess would be that it is
the
> same as Ratio
> - P value and log(error) are some kind of statistics associated with
> the
> hypothesis that intensity 1 is equal to intensity 2
> - It is a two colour array.
>
> I would use Log(Ratio), checking to see what base it is in.
>
> Perhaps you can check out the raw files (which are probably Agilent
> files)
> and figure out what the columns are. They could correspond to
> something like
> gPorcessedSignal/rProcessedSignal
>
> Kasper
>
> On Jun 13, 2008, at 8:12 PM, ss wrote:
>
> Just to follow up a bit, there are 7 corresponding columns for each
> sample.
> For instance, for the 1st array, they are:
>
>
> MBA: US14702370_16012391010920_S01_A01/Log(Ratio)
> MBA: US14702370_16012391010920_S01_A01/Ratio
> MBA: US14702370_16012391010920_S01_A01/Fold Change
> MBA: US14702370_16012391010920_S01_A01/Log(Error)
> MBA: US14702370_16012391010920_S01_A01/P-Value
> MBA: US14702370_16012391010920_S01_A01/Intensity 1
> MBA: US14702370_16012391010920_S01_A01/Intensity 2
>
> I figure I should use Ratio which is Intensity2/Intensity1 for
further
> analysis.
> I really wish I could get started with raw data but I just dont know
> whether
> there is some packages for Agilent data.
>
> Allen
>
> On Fri, Jun 13, 2008 at 11:09 PM, ss <affysnp@gmail.com> wrote:
>
> Thanks Kasper. We downloaded the processed data under the
> accession number 'E-TABM-1' from ArrayExpress at
>
http://www.ebi.ac.uk/microarray-as/aer/#ae-
browse/q=E-TABM-1[2]<http: www.ebi.ac.uk="" microarray-as="" aer="" #ae-="" browse="" q="E-TABM-1%5B2%5D"> >
>
>
> So if you click 'E-TABM-1' and it says that:
>
> Array:Agilent Whole Human Genome Oligo Microarray [G4112A] (» A-
> AGIL-11<http: www.ebi.ac.uk="" microarray-="" as="" aer="" result?queryfor="PhysicalArrayDesign&aAccession=A%2DAGIL%2D11"> >
>
> )
> I guess it is of Agilent format for sure. What does it seem strange
to
> you?
>
> Allen
>
>
>
> On Fri, Jun 13, 2008 at 11:05 PM, Kasper Daniel Hansen <
> khansen@stat.berkeley.edu> wrote:
>
> This does not look at all like an Agilent data file from Feature
> Extractor. I suggest you contact your collaborators and get an idea
> about
> what you should use.
>
> Kasper
>
>
> On Jun 11, 2008, at 4:43 PM, ss wrote:
>
> Dear all,
>
> For whom is familiar with Agilent gene expression data, I would like
> to
> ask help.
>
> We recently received some Agilent gene expression data from our
> collaborators. For individual sample, there are 7 corresponding
> columns:
>
> Unknown:Log(Ratio) Unknown:Ratio Unknown:Fold Change
> Unknown:Log(Error)
> Unknown:P-Value Unknown:Intensity 1 Unknown:Intensity 2
> It seems that Ratio= Intensity1/Intensity2. I wonder whether I
should
> use
> log2(Ratio) or Log(Ratio) or just Ratio for further analysis.
Besides,
> why
> should Ratio be calculated as Intensity1/Intensity2 instead of
> Intensity2/Intensity1?
>
> Thank you very much for your help in advance!
>
> Sincerely,
> Allen
>
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