[BioC] normalizing incomplete arrays [was: limma error message]

Jianping Jin jjin at email.unc.edu
Mon Aug 1 20:41:49 CEST 2005


Thanks Gordon and Naomi for your time and efforts to help me out. The the 
code worked well for me.

One more question. What if I wanted to get the loess normalized expression 
values in addition to M & As? Can I extract the expression values from 
after normalizeWithinArrays or do I have to use something like "loessFit"?

best regards!

JP-

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--On Saturday, July 30, 2005 11:55 PM +1000 Gordon Smyth 
<smyth at wehi.edu.au> wrote:

> As you've deduced, the problem is that your arrays do not a constant
> block size. limma assumes a constant block size for
> normalizeWithinArrays() and imageplot().
>
> Here is a work-around for GenePix data:
>
> for (b in 1:48) {
>          i <- RG$genes$Block==b
>          MA2 <- normalizeWithinArrays(RG[i,],method="loess")
>          if(b==1)
>                  MA <- MA2
>          else
>                  MA <- rbind(MA,MA2)
> }
>
> This will perform print-tip-loess normalization even though there are
> irregular numbers of spots in the print-tip groups.
>
> Gordon
>
>> Date: Fri, 29 Jul 2005 16:42:58 -0400
>> From: Naomi Altman <naomi at stat.psu.edu>
>> Subject: Re: [BioC] limma error message
>> To: Jianping Jin <jjin at email.unc.edu>
>> Cc: Bioconductor at stat.math.ethz.ch
>> Message-ID: <6.2.1.2.2.20050729163707.040c3240 at imap.stat.psu.edu>
>> Content-Type: text/plain; charset="us-ascii"; format=flowed
>>
>> 28*28*12*4
>> [1] 37632
>>
>> But you have 34970  genes.
>>
>> The simplest thing to do is to add the missing lines to your GPR files.
>>
>> You could probably do this in your favorite text manipulation language
>> but with 8 arrays, you might be able to do it manually more quickly.
>>
>> --Naomi
>>
>>
>> At 12:13 PM 7/29/2005, Jianping Jin wrote:
>> > Dear Naomi and BioConductor list,
>> >
>> > Thanks for your kind reply! But I got the same error message after
>> > re-run the program as the following:
>> >
>> >> targets <- readTargets()
>> >> RG <- read.maimages(targets$FileName, source="genepix")
>> > Read 0341_3289_A5_052_HO34K_sample-1_HRef_scan2.gpr
>> > Read 0341_3290_A5_053_HO34K_sample-2_HRef_scan2.gpr
>> > Read 0341_3291_A5_054_HO34K_sample-3_HRef.gpr
>> > Read 0341_3292_A5_055_HO34K_sample-4_HRef.gpr
>> > Read 0341_3293_A5_056_HO34K_sample-5_HRef.gpr
>> > Read 0341_3294_A5_057_HO34K_sample-6_HRef.gpr
>> > Read 0341_3295_A5_058_HO34K_sample-7_HRef.gpr
>> > Read 0341_3296_A5_059_HO34K_sample-8_HRef.gpr
>> >> summary(RG)
>> >        Length Class      Mode
>> > R       279760 -none-     numeric
>> > G       279760 -none-     numeric
>> > Rb      279760 -none-     numeric
>> > Gb      279760 -none-     numeric
>> > targets      1 data.frame list
>> > genes        5 data.frame list
>> >
>> >> dim(RG)
>> > [1] 34970     8
>> >
>> >> getLayout(RG$genes)
>> > $ngrid.r
>> > [1] 12
>> >
>> > $ngrid.c
>> > [1] 4
>> >
>> > $nspot.r
>> > [1] 28
>> >
>> > $nspot.c
>> > [1] 28
>> >
>> > attr(,"class")
>> > [1] "PrintLayout"
>> >
>> >> MA <- normalizeWithinArrays(RG, layout=list(ngrid.r=12, ngrid.c=4,
>> > nspot.r=28, nspot.c=28))
>> > Error: subscript out of bounds
>> >
>> > I checked the data files and found nsopt.r and nspot.c in the last one
>> > third blocks of every files were changed from 28 to 24 and 23
>> > respectively. That may be the reason for the error message. Do you have
>> > any ways to handle this?
>> >
>> > Many thanks!
>> >
>> > JP-
>> >
>> > xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
>> > x Jianping Jin Ph.D.            x
>> > x Bioinformatics scientist      x
>> > x Center for bioinformatics     x
>> > x 3133 Bioinformatics Building  x
>> > x CB# 7104                      x
>> > x University of North Carolina  x
>> > x Chapel Hill, NC 27599         x
>> > x Tel: (919)843-6105            x
>> > x Fax: (919)843-3103            x
>> > x E-mail: jjin at email.unc.edu    x
>> > xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
>> >
>> > --On Thursday, July 28, 2005 5:17 PM -0400 Naomi Altman
>> > <naomi at stat.psu.edu> wrote:
>> >
>> >> layout=list(ngrid.r= ,ngrid.c= ,nspot.r= , nspot.c= )
>> >>
>> >> Your array is laid out in blocks according to the print tip
>> >> configuration of the printer.
>> >> nspot.r is the number of rows in each block
>> >> nspot.c is the number of columns in each block
>> >>
>> >> ngrid.r is the number of rows of blocks
>> >> ngrid.c is the number of columns of blocks
>> >>
>> >> e.g. a typical 8 tip design might have 4 rows and 2 columns of blocks
>> >>
>> >> If you don't know the right numbers to put in, look at the raw data -
>> >> the block number and row and column are output to your expression
>> >> file.
>> >>
>> >> --Naomi
>> >>
>> >> At 04:49 PM 7/28/2005, you wrote:
>> >>> Thanks Naomi! Could you explain how to specify parameter "layout"?
>> >>> JP-
>> >>>
>> >>> --On Thursday, July 28, 2005 3:35 PM -0400 Naomi Altman
>> >>> <naomi at stat.psu.edu> wrote:
>> >>>
>> >>>> I got this message when I used the wrong layout file.
>> >>>>
>> >>>> --Naomi
>> >>>>
>> >>>> At 03:03 PM 7/28/2005, Jianping Jin wrote:
>> >>>>> Dear BioConductor list,
>> >>>>>
>> >>>>> I got an error, Error: subscipt out of bounds, when I started to
>> >>>>> run a limma program normalizeWithinArrays(). What I wanted to do
>> >>>>> was to normalize eight cDNA slides. >dim(RG): 34970      8.
>> >>>>>
>> >>>>> I used >debug(normalizeWithinArrays) and then
>> >>>>> > normalizeWithinArrays(RG) to debug by either Browse[1]> n or c as
>> >>>>> well. But when I retried the program it gave me the same error
>> >>>>> message above.
>> >>>>>
>> >>>>> I will highly appreciate it if any one could give it shot.
>> >>>>>
>> >>>>>
>> >>>>> Best regards!
>> >>>>>
>> >>>>> JP-
>> >>>>>
>> >>>>> xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
>> >>>>> x Jianping Jin Ph.D.            x
>> >>>>> x Bioinformatics scientist      x
>> >>>>> x Center for bioinformatics     x
>> >>>>> x 3133 Bioinformatics Building  x
>> >>>>> x CB# 7104                      x
>> >>>>> x University of North Carolina  x
>> >>>>> x Chapel Hill, NC 27599         x
>> >>>>> x Tel: (919)843-6105            x
>> >>>>> x Fax: (919)843-3103            x
>> >>>>> x E-mail: jjin at email.unc.edu    x
>> >>>>> xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
>> >>>>>
>> >>>>
>> >>>> Naomi S. Altman                                814-865-3791 (voice)
>> >>>> Associate Professor
>> >>>> Bioinformatics Consulting Center
>> >>>> Dept. of Statistics                              814-863-7114 (fax)
>> >>>> Penn State University                         814-865-1348
>> >>>> (Statistics) University Park, PA 16802-2111
>



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