question about scale factor for Affy chip
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@james-anderson-1641
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Guido Hooiveld ★ 4.1k
@guido-hooiveld-2020
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Wageningen University, Wageningen, the …
It is A, since you scale all arrays to the same target value (100 is default in BioC, 500 is default in GCOS). According to Affymetrix'criteria the difference between the SFs should not vary more than a factor of 3. Since in your case the SFs are within the same range (10.-10.2), the SF does not indicate a problem. If you would have had 10.2. 10.1, 10.4, 40.9 and 2.7, the SF indiactes the last two arrays deviate from the other arrays. HTH, Guido > -----Original Message----- > From: bioconductor-bounces at stat.math.ethz.ch > [mailto:bioconductor-bounces at stat.math.ethz.ch] On Behalf Of > James Anderson > Sent: 29 June 2007 16:32 > To: bioconductor > Subject: [BioC] question about scale factor for Affy chip > > Hi, > > I am using the R package SimpleAffy, now I have a conceptual > question regarding the scale factor. Seems that Affymetrix > recommend the scale factor should be within 3 fold, the way I > understand the scale factor is that it is the ratio between > the trimmed mean of one chip (normalized by MAS5) to the > target value (say, 100 or 500). According to the manual for > QC in SimpleAffy: > > "Large variations in scale factors signal cases where the > normalisation assumptions are likely to fail due to issues > with sample quality or amount of starting material." > > I am getting a little confused about EXACTLY what does scale > factor mean: > A. The ratio between the trimmed mean of each chip to a > target value B. The ratio between the trimmed mean of each > chip to the trimmed mean of all the chips. > > because the above section seems to me that as long as there > is small variation in the scale factor, the quality is no > problem. What about all the arrays have scale factor in the > range (10.1-10.2)? The scale factor has almost no variation, > but they are much higher than 3. or I am misunderstanding some part? > > > Thanks a lot! > > James > > > --------------------------------- > Pinpoint customers who are looking for what you sell. > [[alternative HTML version deleted]] > > _______________________________________________ > Bioconductor mailing list > Bioconductor at stat.math.ethz.ch > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: > http://news.gmane.org/gmane.science.biology.informatics.conductor >
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Yep, but plz note that for the calculation of the SF no reference array is used, but just a set value of 100 (500). This is in contrast to e.g. dChip, in which of the [experimental] arrays is used as reference, and therefore the set value varies between experiments. > -----Original Message----- > From: bioconductor-bounces at stat.math.ethz.ch > [mailto:bioconductor-bounces at stat.math.ethz.ch] On Behalf Of > James Anderson > Sent: 29 June 2007 17:40 > To: Hooiveld, Guido; bioconductor > Subject: Re: [BioC] question about scale factor for Affy chip > > Thanks for your email. So the criterion "SFs should be within > 3 fold" actually means that the ratio of the scale factor of > each array to the scale factor of the reference (mean) array > should be within 3 fold. > > "Hooiveld, Guido" <guido.hooiveld at="" wur.nl=""> wrote: > It is A, since you scale all arrays to the same target value > (100 is default in BioC, 500 is default in GCOS). > > According to Affymetrix'criteria the difference between the > SFs should not vary more than a factor of 3. Since in your > case the SFs are within the same range (10.-10.2), the SF > does not indicate a problem. If you would have had 10.2. > 10.1, 10.4, 40.9 and 2.7, the SF indiactes the last two > arrays deviate from the other arrays. > > HTH, > Guido > > > > -----Original Message----- > > From: bioconductor-bounces at stat.math.ethz.ch > > [mailto:bioconductor-bounces at stat.math.ethz.ch] On Behalf Of James > > Anderson > > Sent: 29 June 2007 16:32 > > To: bioconductor > > Subject: [BioC] question about scale factor for Affy chip > > > > Hi, > > > > I am using the R package SimpleAffy, now I have a > conceptual question > > regarding the scale factor. Seems that Affymetrix recommend > the scale > > factor should be within 3 fold, the way I understand the > scale factor > > is that it is the ratio between the trimmed mean of one chip > > (normalized by MAS5) to the target value (say, 100 or 500). > According > > to the manual for QC in SimpleAffy: > > > > "Large variations in scale factors signal cases where the > > normalisation assumptions are likely to fail due to issues > with sample > > quality or amount of starting material." > > > > I am getting a little confused about EXACTLY what does scale factor > > mean: > > A. The ratio between the trimmed mean of each chip to a > target value > > B. The ratio between the trimmed mean of each chip to the > trimmed mean > > of all the chips. > > > > because the above section seems to me that as long as there > is small > > variation in the scale factor, the quality is no problem. > What about > > all the arrays have scale factor in the range (10.1-10.2)? > The scale > > factor has almost no variation, but they are much higher > than 3. or I > > am misunderstanding some part? > > > > > > Thanks a lot! > > > > James > > > > > > --------------------------------- > > Pinpoint customers who are looking for what you sell. > > [[alternative HTML version deleted]] > > > > _______________________________________________ > > Bioconductor mailing list > > Bioconductor at stat.math.ethz.ch > > https://stat.ethz.ch/mailman/listinfo/bioconductor > > Search the archives: > > http://news.gmane.org/gmane.science.biology.informatics.conductor > > > > > > --------------------------------- > > [[alternative HTML version deleted]] > > _______________________________________________ > Bioconductor mailing list > Bioconductor at stat.math.ethz.ch > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: > http://news.gmane.org/gmane.science.biology.informatics.conductor >
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