Hi,
I'm trying to normalize between arrays using the limmma package so I
can run SAM. I want to know whether we can read log2 ratio data into
R? I'm using Agilent's extraction program to extract cDNA arrays
data and the image analysis program from Agilent automatically
normalize within array, giving the normalized intensity and log
ratio. Is there anyway I can by-pass reading Mean/Median foreground
and background intensity and directly feed the processed (background
subtracted and normalized) intensity into R?
Another question: Can I also input a column that filters spots which
were unsatisfactory according to the image analysis program? And how
can I can remove these bad spots from further calculation in R using
the limma package or any other functions in R?
Thanks,
Anna
Anna,
Agilent's extraction software also provides a flat file with
all the meaurements, median and mean background and foregounds,
so you can use those as an input,
I found this very amenable to use of Bioconductor
in what I do,
Best
Susan Holmes
Associate Professor
Statistics
On Wed, 10 Dec 2003, Anna Cao wrote:
> Hi,
>
> I'm trying to normalize between arrays using the limmma package so I
> can run SAM. I want to know whether we can read log2 ratio data into
> R? I'm using Agilent's extraction program to extract cDNA arrays
> data and the image analysis program from Agilent automatically
> normalize within array, giving the normalized intensity and log
> ratio. Is there anyway I can by-pass reading Mean/Median foreground
> and background intensity and directly feed the processed (background
> subtracted and normalized) intensity into R?
>
> Another question: Can I also input a column that filters spots which
> were unsatisfactory according to the image analysis program? And how
> can I can remove these bad spots from further calculation in R using
> the limma package or any other functions in R?
>
> Thanks,
>
> Anna
>
> _______________________________________________
> Bioconductor mailing list
> Bioconductor@stat.math.ethz.ch
> https://www.stat.math.ethz.ch/mailman/listinfo/bioconductor
>
At 06:15 AM 11/12/2003, Anna Cao wrote:
>Hi,
>
>I'm trying to normalize between arrays using the limmma package so I
>can run SAM. I want to know whether we can read log2 ratio data into
>R? I'm using Agilent's extraction program to extract cDNA arrays
>data and the image analysis program from Agilent automatically
>normalize within array, giving the normalized intensity and log
>ratio. Is there anyway I can by-pass reading Mean/Median foreground
>and background intensity and directly feed the processed (background
>subtracted and normalized) intensity into R?
This is easy in R. Use the Agilent flat file, use read.table(), then
collect the normalized columns into a matrix in R. The matrix can then
but
used as input for example to the lmFit() command in limma. You'll have
to
do the input yourself though, there aren't any canned limma commands
to do
it. If you need help with this, you might want to ask for help on the
R-help mailing list.
>Another question: Can I also input a column that filters spots which
>were unsatisfactory according to the image analysis program? And how
>can I can remove these bad spots from further calculation in R using
>the limma package or any other functions in R?
Using limma, you set the appropriate elements of the 'weights' matrix
to
zero for those spots. This applies to both normalization and linear
modelling functions in limma. Normally the weights matrix is set
automatically using the wt.fun argument of read.maimages, but you will
need
to create it yourself if you're reading in the data manually as it
were.
Gordon
>Thanks,
>
>Anna
Progress update (summarized from my forum for such matters at
http://www.theschedule.net/forum/gforum.cgi?forum=20&do=forum_view):
Briefly, I created a four-node cluster out of Pentium-III boxes and
Debian Linux/openMosix. I saw no significant performance boost of
ReadAffy or expresso using the set of 165 .CEL files from Harvard.
None
of the processes migrated, as they say in the world of high-
performance
computing. R.bin runs in one process, and everything it does seems to
stay in that process. No real opportunity for parallelization here,
at
least not on openMosix.
I'd like to analyze these chips in a reasonable amount of time,
without
paying Dell $45,000 for 4-Xeon SMP server.
I worry what we'll do with 1,000 .CEL files. The analytical
techniques
work well, but pretty slow even if your amp "goes to 11."
Any thoughts?
Michael Benjamin, MD
Emory University
Hi,
I'm able to read the Agilent flat file using the read.table function
but
what if I have a large amount of arrays to input. This function seems
to
only input one table at a time. Are there any ways to read in multiple
files
simultaneously (as in the read.maimages function)? Can you please list
the
commands/functions I would need to perform the reading in normalized
signals
task? Then I can go and look through help in R for them?
How would I go about defining the weight matrix for each array since
the
source for the Agilent analysis program is not included in the
read.maimages
function? I have a column of filter values in the Agilent txt file
which was
read in through read.maimages (manually through the column argument).
How
can I define this filter column?
Thanks for you time,
Anna
-----Original Message-----
From: Gordon Smyth [mailto:smyth@wehi.edu.au]
Sent: Wednesday, December 10, 2003 4:45 PM
To: Anna Cao
Cc: bioconductor@stat.math.ethz.ch
Subject: Re: [BioC] limma package
At 06:15 AM 11/12/2003, Anna Cao wrote:
>Hi,
>
>I'm trying to normalize between arrays using the limmma package so I
>can run SAM. I want to know whether we can read log2 ratio data into
>R? I'm using Agilent's extraction program to extract cDNA arrays
>data and the image analysis program from Agilent automatically
>normalize within array, giving the normalized intensity and log
>ratio. Is there anyway I can by-pass reading Mean/Median foreground
>and background intensity and directly feed the processed (background
>subtracted and normalized) intensity into R?
This is easy in R. Use the Agilent flat file, use read.table(), then
collect the normalized columns into a matrix in R. The matrix can then
but
used as input for example to the lmFit() command in limma. You'll have
to
do the input yourself though, there aren't any canned limma commands
to do
it. If you need help with this, you might want to ask for help on the
R-help mailing list.
>Another question: Can I also input a column that filters spots which
>were unsatisfactory according to the image analysis program? And how
>can I can remove these bad spots from further calculation in R using
>the limma package or any other functions in R?
Using limma, you set the appropriate elements of the 'weights' matrix
to
zero for those spots. This applies to both normalization and linear
modelling functions in limma. Normally the weights matrix is set
automatically using the wt.fun argument of read.maimages, but you will
need
to create it yourself if you're reading in the data manually as it
were.
Gordon
>Thanks,
>
>Anna