Dear Dr. Laurent Gautier,
Thanks for your kind reply.
> Quantile normalization is usually performed on untransformed data
("raw-scale").
I read some journals on RMA or RMA++, and I made sure that raw-scale
is
usually used for quantile normarlization as you say.
> Missing values are just ignored and left as such (missing values).
I'm afraid I don't quite understand what you say on how to treat the
N/
A.
Based on the program written in "normalizeBetweenArrays", it seems
that the
N/A is first replaced with the median, but after ranking, averaging
and
re-ordering to the original position, it's transformed back to N/A.
Is
this correct?
I described the example herein:
step 1: Calculating median in each column
> ngenes <- 5
> narrays <- 2
> x <- matrix(c(1:10),ngenes,narrays)
> x[2,1] <- x[3,1] <- NA
> x[4,2] <- x[5,2] <- NA
> x
[,1] [,2]
[1,] 1 6
[2,] NA 7
[3,] NA 8
[4,] 4 NA
[5,] 5 NA
> (xm <- apply(x,2,median,na.rm=T))
[1] 4 7
step 2: Replacing N/A in each column with median
> x[2,1] <- x[3,1] <- xm[1]
> x[4,2] <- x[5,2] <- xm[2]
[,1] [,2]
[1,] 1 6
[2,] 4 7
[3,] 4 8
[4,] 4 7
[5,] 5 7
step 3: Sorting values in each column in descending order
[,1] [,2]
[1,] 5 8
[2,] 4 7
[3,] 4 7
[4,] 4 7
[5,] 1 6
step 4: Averaging values in each rank
[,1] [,2] Average
[1,] 5 8 6.5
[2,] 4 7 5.5
[3,] 4 7 5.5
[4,] 4 7 5.5
[5,] 1 6 3.5
step 5: Replacing the values in each column with the average
[,1] [,2] Average
[1,] 6.5 6.5 6.5
[2,] 5.5 5.5 5.5
[3,] 5.5 5.5 5.5
[4,] 5.5 5.5 5.5
[5,] 3.5 3.5 3.5
step 6: Re-sorting the values in each column at original positions
[,1] [,2]
[1,] 3.5 3.5
[2,] 5.5 5.5
[3,] 5.5 6.5
[4,] 5.5 5.5
[5,] 6.5 5.5
step 7: Replacing the values with N/A at original positions
[,1] [,2]
[1,] 3.5 3.5
[2,] NA 5.5
[3,] NA 6.5
[4,] 5.5 NA
[5,] 6.5 NA
This result corresponds to normalizeBetweenArrays() result;
> x <- matrix(c(1:10),ngenes,narrays)
> x[2,1] <- x[3,1] <- NA
> x[4,2] <- x[5,2] <- NA
> (y <- normalizeBetweenArrays(x))
[,1] [,2]
[1,] 3.5 3.5
[2,] NA 5.5
[3,] NA 6.5
[4,] 5.5 NA
[5,] 6.5 NA
J, K
--- On Mon, 2011/8/1, Laurent Gautier <laurent at="" cbs.dtu.dk=""> wrote:
On 2011-08-01 06:55, qwertyui_period at yahoo.co.jp wrote:
> Dear all,
>
> My environment is limma Version 3.2.2, R version 2.10.1, and
Windows XP.
> I'm going to normalize the microarray data by
"normalizeBetweenArrays"
> which is the quantile normalization function in "limma" package.
> I have read the "usersguide.pdf" in bioconductor website, however,
I still
> have two questions.
>
> Question 1: Which is proper to use for quantile normalization: raw-
scale or
> log2-scale values ?
> The quantile normalization includes the step of calculating
arithmetic
> mean,
> so I suppose the raw-scale values should be used, though the
microarray
> data is generally log2-scale values.
Quantile normalization is usually performed on untransformed data
("raw-scale").
Log2 transformation comes after (before probe summary when using RMA
or
RMA-like approaches).
> Question 2: How does "normalizeBetweenArrays" deal $B!H(BN/A$B!I(B
in data ?
Missing values are just ignored and left as such (missing values).
Hoping this helps,
L.
> Example code 1,
>
>> ngenes<- 3
>> narrays<- 2
>> x<- matrix(c(3,1,5,6,4,2),ngenes,narrays)
> [,1] [,2]
> [1,] 3 6
> [2,] 1 4
> [3,] 5 2
>
>> (y<- normalizeBetweenArrays(x))
> [,1] [,2]
> [1,] 3.5 5.5
> [2,] 1.5 3.5
> [3,] 5.5 1.5
>
> I understand the process of "normalizeBetweenArrays" is devided
into 4
> steps as follows:
>
> step 1: Sorting values in each column in descending order
>
> [,1] [,2]
> [1,] 5 6
> [2,] 3 4
> [3,] 1 2
>
> step 2: Averaging values in each rank
>
> [,1] [,2] Average
> [1,] 5 6 5.5
> [2,] 3 4 3.5
> [3,] 1 2 1.5
>
> step 3: Replacing the values in each column with the average
>
> [,1] [,2] Average
> [1,] 5.5 5.5 5.5
> [2,] 3.5 3.5 3.5
> [3,] 1.5 1.5 1.5
>
> step 4: Re-sorting the values in each column at original positions
>
> [,1] [,2]
> [1,] 3.5 5.5
> [2,] 1.5 3.5
> [3,] 5.5 1.5
>
> Then, how does "normalizeBetweenArrays" deal $B!H(BN/A$B!I(B in
data ?
>
> Example code 2,
>
>> (x<- matrix(c(NA,1,5,6,4,2),ngenes,narrays))
> [,1] [,2]
> [1,] NA 6
> [2,] 1 4
> [3,] 5 2
>
> (y<- normalizeBetweenArrays(x))
>
> [,1] [,2]
> [1,] NA 5.5
> [2,] 1.5 3.5
> [3,] 5.5 1.5
>
> Thanks in advance !
>
>
> [[alternative HTML version deleted]]
>
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