Description Usage Arguments Details Value Note Author(s) References See Also Examples
Corrects the gene- and slide specific dye bias in a data set, using the GASSCO method by Margaritis et al.
1 2 3 4 |
data.norm |
A |
iGSDBs |
A data frame with the intrinsic gene specific dye bias per reporter
(i.e., oligo or cDNA). The data frame would typically have come
from a call to The data frame must have (at least) the following columns:
The order of the rows in this data frame is irrelevant. There must
be no rows with duplicate For any reporter in |
estimator.subset |
An index indicating which reporters are fit to be used as estimators of the slide bias. This set of reporters is used throughout the whole data set. Reporters that are typically excluded are those corresponding to parasitic DNA elements or mitochondrial genes. |
application.subset |
An index indicating which values must be dye
bias-corrected. It should be either a vector with as many values as
spots, or a matrix of the same dimensions as
Often it is prudent not to dye bias-correct measurements that are
close to the detection limit or close to signal saturation. A
convenience function for this is provided; see |
dyebias.percentile |
The slide bias estimation uses a small subset of reporters having the strongest green or red iGSDB, as specified by this percentile. The default should suffice in practically all cases. |
minmaxA.perc |
To obtain a robust estimate of the slide bias, the range of the
average expression A is trimmed by |
minA.abs |
If specified, reporters with an average expression
(A) lower than this value are never considered as estimators
of the slide bias. If not specified, reporters with an
A-percentile < |
maxA.abs |
If specified, reporters with an average expression
(A) greater than this are never considered as estimators of the
slide bias. If not specified, reporters with an A-percentile <
|
verbose |
Logical speficying whether to be verbose or not |
This function corrects the gene-specific dye bias of two-colour microarrays with the GASSCO method. This method is general, robust and fast, and is based on the observation that the total bias per gene is the product of a slide-specific factor (strongly related to the labeling percentage) and an intrinsic gene-specific factor (iGSDB), which is strongly related to the probe sequence.
The slide bias is estimated from the total bias of the
dyebias.percentile
percentage of reporters having the strongest
iGSDB. The iGSDBs can be estimated with
dyebias.estimate.iGSDBs
.
If the signal of certain oligos is too weak, or in contrast, tends to
be saturated, they are no good estimator of the slide bias.
Therefore, only reporters with an average expression level A
that is not too extreme are allowed to be slide bias estimators. (This
is the reason for the A
-column in the iGSDBs
data
frame).
Full control over which reporters to allow as slide bias estimators is
given by the arguments minmaxA.perc
, minA.abs
, and
maxA.abs
; see there for details. To not exclude any reporter
(e.g., when A is not available and therefore artificially set),
you can use minA.abs= -Inf
and maxA.abs = Inf
.
For further details concerning the method, see the dyebias
vignette and the publication. If your research benefits from using this
package, we kindly request that you cite this work.
The data returned is a list wit the following elements
data.corrected |
A |
estimators |
Another list, containing the details of the
reporters that were used to obtain an estimate of the slide bias.
The contents of the
|
summary |
A data frame summarizing the correction process per slide. It consist of the following columns:
|
data.uncorrected |
The uncorrected input |
Note that the input data should be normalized, and that the dye swaps should not have been swapped back (if needed, this can of course be done afterwards).
Philip Lijnzaad p.lijnzaad@umcutrecht.nl
Margaritis, T., Lijnzaad, P., van Leenen, D., Bouwmeester, D., Kemmeren, P., van Hooff, S.R and Holstege, F.C.P. (2009). Adaptable gene-specific dye bias correction for two-channel DNA microarrays. Molecular Systems Biology, 5:266, 2009. doi: 10.1038/msb.2009.21.
dyebias.estimate.iGSDBs
,
dyebias.application.subset
,
dyebias.rgplot
,
dyebias.maplot
,
dyebias.boxplot
,
dyebias.trendplot
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 | ## First load data and estimate the iGSDBs
## (see dyebias.estimate.iGSDBs)
### choose the estimators and which spots to correct:
estimator.subset <- dyebias.umcu.proper.estimators(maInfo(maGnames(data.norm)))
### choose which genes to dye bias correct:
application.subset <- (maW(data.norm) == 1 &
dyebias.application.subset(data.raw=data.raw, use.background=TRUE))
### do the correction:
correction <- dyebias.apply.correction(data.norm=data.norm,
iGSDBs = iGSDBs.estimated,
estimator.subset=estimator.subset,
application.subset = application.subset,
verbose=FALSE)
## Not run:
edit(correction$summary)
## End(Not run)
## give overview:
correction$summary[,c("slide", "file", "avg.correction", "reduction.perc", "p.value")]
## and summary:
summary(as.numeric(correction$summary[, "reduction.perc"]))
|
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