Hi,
I apologize for this being off-topic- it's really a statistical
question
but I'd be interested in the community's input. If I run a 'per gene'
2-way
ANOVA on single channel microarray data (i.e., each gene is tested
separately by 2-Way ANOVA), should I run multiple testing correction
for
each factor and interaction separately? Alternatively, should I use an
overall (omnibus) F-test, correct that for multiple testing, and treat
the
main effects and interaction results as post-hoc to the overall test?
Thanks,
-E
Eric Blalock, PhD
Dept Pharmacology, UKMC
859 323-8033
STATEMENT OF CONFIDENTIALITY\ \ The contents of this e-mail
...{{dropped}}
I am absolutely no expert in multiple comparison / multiple testing /
gene
expression data analysis, so take the following with appropriate dose
of
salt:
It really depends on what you are looking to get out of the data.
Just
because you have multi-factor data with > 2 levels and thousands of
responses, it doesn't automatically mean that the usual multiple
comparison
procedures are appropriate. You design the experiment to answer some
specific questions (hopefully). How you analyze the data depends
greatly on
what those questions are, and (hopefully, therefore) how the
experiment is
designed.
Best,
Andy
> From: Eric
>
> Hi,
>
> I apologize for this being off-topic- it's really a
> statistical question
> but I'd be interested in the community's input. If I run a
> 'per gene' 2-way
> ANOVA on single channel microarray data (i.e., each gene is tested
> separately by 2-Way ANOVA), should I run multiple testing
> correction for
> each factor and interaction separately? Alternatively, should
> I use an
> overall (omnibus) F-test, correct that for multiple testing,
> and treat the
> main effects and interaction results as post-hoc to the overall
test?
>
> Thanks,
> -E
>
> Eric Blalock, PhD
> Dept Pharmacology, UKMC
> 859 323-8033
>
> STATEMENT OF CONFIDENTIALITY\ \ The contents of this e-mail
> ...{{dropped}}
>
> _______________________________________________
> Bioconductor mailing list
> Bioconductor@stat.math.ethz.ch
> https://www.stat.math.ethz.ch/mailman/listinfo/bioconductor
>
>
Hi Andy,
Thanks for the reply. My reasoning here is a little Byzantine so bear
with me.
If the significant results are relatively evenly distributed across
the
main effects and the interaction (about the same number of genes found
in
each), then using the omnibus test will not make much of a difference.
However, say one of the two main effects is much stronger than the
other,
then I have a case where the overall test will pick up all of those
changes
from the 'powerful' treatment (or most of them). Because of that,
multiple
testing correction at the overall level will allow genes with larger
p-values from the second main effect through the filter compared to
the
list of genes that would make it through a multiple testing correction
applied at the level of the second main effect.
Contrast this with the case where multiple testing is applied
separately to
each of the three outputs. Here the first main effect is relatively
unaffected, but the second main effect is nuked (if the second main
effect
has no more genes than would be expected by chance). IMHO it doesn't
matter
what the original question was, the two multiple testing corrections
change
the list of genes and the experimental question does not address which
of
these procedures should be used. It would be disingenuous to say
"Well,
we're mainly interested in main effect 2 (the weak one), so we'll use
the
overall correction and at least see a list of genes" or "We wanted to
disagree with previous work about main effect two's importance to
research
so we used individual correction to show the world that main effect
two is
not doing anything". Perhaps the proportion of genes assigned an
interaction significance could be used to gauge the dependence of the
two
main effects; the more dependent they are, the more applicable the
overall
testing correction. While the smaller the proportion of genes showing
an
interaction term, the more appropriate independent correction for each
main
effect would be.
At 03:05 PM 7/27/2004, you wrote:
>I am absolutely no expert in multiple comparison / multiple testing /
gene
>expression data analysis, so take the following with appropriate dose
of
>salt:
>
>It really depends on what you are looking to get out of the data.
Just
>because you have multi-factor data with > 2 levels and thousands of
>responses, it doesn't automatically mean that the usual multiple
comparison
>procedures are appropriate. You design the experiment to answer some
>specific questions (hopefully). How you analyze the data depends
greatly on
>what those questions are, and (hopefully, therefore) how the
experiment is
>designed.
>
>Best,
>Andy
>
> > From: Eric
> >
> > Hi,
> >
> > I apologize for this being off-topic- it's really a
> > statistical question
> > but I'd be interested in the community's input. If I run a
> > 'per gene' 2-way
> > ANOVA on single channel microarray data (i.e., each gene is tested
> > separately by 2-Way ANOVA), should I run multiple testing
> > correction for
> > each factor and interaction separately? Alternatively, should
> > I use an
> > overall (omnibus) F-test, correct that for multiple testing,
> > and treat the
> > main effects and interaction results as post-hoc to the overall
test?
> >
> > Thanks,
> > -E
> >
> > Eric Blalock, PhD
> > Dept Pharmacology, UKMC
> > 859 323-8033
> >
> > STATEMENT OF CONFIDENTIALITY\ \ The contents of this e-mail
> > ...{{dropped}}
> >
> > _______________________________________________
> > Bioconductor mailing list
> > Bioconductor@stat.math.ethz.ch
> > https://www.stat.math.ethz.ch/mailman/listinfo/bioconductor
> >
> >
>
>
>---------------------------------------------------------------------
---------
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>information of Merck & Co., Inc. (One Merck Drive, Whitehouse
Station, New
>Jersey, USA 08889), and/or its affiliates (which may be known outside
the
>United States as Merck Frosst, Merck Sharp & Dohme or MSD and in
Japan, as
>Banyu) that may be confidential, proprietary copyrighted and/or
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>privileged. It is intended solely for the use of the individual or
entity
>named on this message. If you are not the intended recipient, and
have
>received this message in error, please notify us immediately by reply
>e-mail and then delete it from your system.
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---------
Eric Blalock, PhD
Dept Pharmacology, UKMC
859 323-8033
STATEMENT OF CONFIDENTIALITY\ \ The contents of this e-mail
...{{dropped}}
I think I asked this question on the list before but regarding one-way
ANOVA and pairwise comparison. And I am no expert in multiple
comparison
either.
In the following paper, there are two main effect group and time. If I
remember correctly the authors argue that interaction term is most
important (otherwise one-way ANOVA would suffice) followed by groups
effect.
Statistical tests for identifying differentially expressed genes in
time-course microarray experiments.
Park T., Yi S.G., Lee S., Lee S.Y., Yoo D.H., Ahn J.I., Lee Y.S.
Bioinformatics 2003; 19(6):694-703
12691981
(Don't you just hate p-values ?)
On Tue, 2004-07-27 at 21:38, Eric wrote:
> Hi Andy,
>
> Thanks for the reply. My reasoning here is a little Byzantine so
bear with me.
>
> If the significant results are relatively evenly distributed across
the
> main effects and the interaction (about the same number of genes
found in
> each), then using the omnibus test will not make much of a
difference.
> However, say one of the two main effects is much stronger than the
other,
> then I have a case where the overall test will pick up all of those
changes
> from the 'powerful' treatment (or most of them). Because of that,
multiple
> testing correction at the overall level will allow genes with larger
> p-values from the second main effect through the filter compared to
the
> list of genes that would make it through a multiple testing
correction
> applied at the level of the second main effect.
>
> Contrast this with the case where multiple testing is applied
separately to
> each of the three outputs. Here the first main effect is relatively
> unaffected, but the second main effect is nuked (if the second main
effect
> has no more genes than would be expected by chance). IMHO it doesn't
matter
> what the original question was, the two multiple testing corrections
change
> the list of genes and the experimental question does not address
which of
> these procedures should be used. It would be disingenuous to say
"Well,
> we're mainly interested in main effect 2 (the weak one), so we'll
use the
> overall correction and at least see a list of genes" or "We wanted
to
> disagree with previous work about main effect two's importance to
research
> so we used individual correction to show the world that main effect
two is
> not doing anything". Perhaps the proportion of genes assigned an
> interaction significance could be used to gauge the dependence of
the two
> main effects; the more dependent they are, the more applicable the
overall
> testing correction. While the smaller the proportion of genes
showing an
> interaction term, the more appropriate independent correction for
each main
> effect would be.
>
>
> At 03:05 PM 7/27/2004, you wrote:
> >I am absolutely no expert in multiple comparison / multiple testing
/ gene
> >expression data analysis, so take the following with appropriate
dose of
> >salt:
> >
> >It really depends on what you are looking to get out of the data.
Just
> >because you have multi-factor data with > 2 levels and thousands of
> >responses, it doesn't automatically mean that the usual multiple
comparison
> >procedures are appropriate. You design the experiment to answer
some
> >specific questions (hopefully). How you analyze the data depends
greatly on
> >what those questions are, and (hopefully, therefore) how the
experiment is
> >designed.
> >
> >Best,
> >Andy
> >
> > > From: Eric
> > >
> > > Hi,
> > >
> > > I apologize for this being off-topic- it's really a
> > > statistical question
> > > but I'd be interested in the community's input. If I run a
> > > 'per gene' 2-way
> > > ANOVA on single channel microarray data (i.e., each gene is
tested
> > > separately by 2-Way ANOVA), should I run multiple testing
> > > correction for
> > > each factor and interaction separately? Alternatively, should
> > > I use an
> > > overall (omnibus) F-test, correct that for multiple testing,
> > > and treat the
> > > main effects and interaction results as post-hoc to the overall
test?
> > >
> > > Thanks,
> > > -E
> > >
> > > Eric Blalock, PhD
> > > Dept Pharmacology, UKMC
> > > 859 323-8033
> > >
> > > STATEMENT OF CONFIDENTIALITY\ \ The contents of this e-mail
> > > ...{{dropped}}
> > >
> > > _______________________________________________
> > > Bioconductor mailing list
> > > Bioconductor@stat.math.ethz.ch
> > > https://www.stat.math.ethz.ch/mailman/listinfo/bioconductor
> > >
> > >
> >
> >
> >-------------------------------------------------------------------
-----------
> >Notice: This e-mail message, together with any attachments,
contains
> >information of Merck & Co., Inc. (One Merck Drive, Whitehouse
Station, New
> >Jersey, USA 08889), and/or its affiliates (which may be known
outside the
> >United States as Merck Frosst, Merck Sharp & Dohme or MSD and in
Japan, as
> >Banyu) that may be confidential, proprietary copyrighted and/or
legally
> >privileged. It is intended solely for the use of the individual or
entity
> >named on this message. If you are not the intended recipient, and
have
> >received this message in error, please notify us immediately by
reply
> >e-mail and then delete it from your system.
> >-------------------------------------------------------------------
-----------
>
> Eric Blalock, PhD
> Dept Pharmacology, UKMC
> 859 323-8033
>
> STATEMENT OF CONFIDENTIALITY\ \ The contents of this e-mail
...{{dropped}}
>
> _______________________________________________
> Bioconductor mailing list
> Bioconductor@stat.math.ethz.ch
> https://www.stat.math.ethz.ch/mailman/listinfo/bioconductor
>