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Erika Melissari
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250
@erika-melissari-2798
Last seen 10.2 years ago
Dear all,
I am starting a statistical analysis for a new experiment and I have
some doubts about quality control filtering procedure.
I know quality control is essential in order to obtain a "reliable"
result at the end of statistical analysis.
Usually I discard spots by setting a convenient wt.fun as argument of
read.maimages: by this function I assign a weight 0 to each spot with
low SNR, or high percentage of saturated pixels, or GenePix "bad"
flag, or simlpy if the spot is a ControlSpot (I do not use control
spot for normalization process).
I read the array quality method of Ritchie et al (2006) implemented in
LIMMA. If I do not make a mistake, this method assign an "array
weight" to arrays with poor quality (reproducibility) and use it to
down-weight their observations. arrayWeights() produces the new array
of weights used in lmFit().
I am excited about using this method for my next analysis, but I do
not manage to understand how I can put together my personal quality
filtering procedure (realized by wt.fun), wich produces an array of
weights, and the weights obtained by arrayWeights().
Firstly, is it right putting together the spot weights with array
weights?
Are the information regarding saturation or low SNR taked into account
from arrayWeights() method?
Is it essential, in your opinion, using these last two charateristics
to discard spots?
In my last experiment I discard a lot of spot (about 60%) due to low
SNR and I am a bit worried about fitting a model with a so poor number
of survived spots....
Or more simply, should I use only arrayWeights() and Spot Type File to
eliminate control spots from lmFit?
Thank you for any help
Erika
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