how to rank affy probesets by their probe-effect magnitude
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Robert Castelo ★ 3.4k
@rcastelo
Last seen 2 hours ago
Barcelona/Universitat Pompeu Fabra
dear list, i'm searching for a way to rank affy probesets from classical 3' affy arrays by their probe effect magnitude. i mean that i would like to know if a probeset is has a larger probe-specific effect than another one. i guess the solution should be in the affyPLM package since if i do library(affy) library(affyPLM) ab <- ReadAffy() pset <- fitPLM(ab) i obtain an object (pset) of the PLMset class which contains slots 'probe.coefs' and 'se.probe.coefs', where each is a list as many keys as probesets and where each probeset contains information on the probe effect of each probe within the probeset: head(names(pset at probe.coefs)) [1] "1000_at" "1001_at" "1002_f_at" "1003_s_at" "1004_at" "1005_at" head(names(pset at se.probe.coefs)) [1] "1000_at" "1001_at" "1002_f_at" "1003_s_at" "1004_at" "1005_at" pset at probe.coefs[[1]] Overall probe_1 0.97287528 probe_2 0.61454806 probe_3 -2.81701693 probe_4 1.68063395 probe_5 -3.31991235 probe_6 1.56657388 probe_7 -3.30256264 probe_8 -1.99431231 probe_9 -0.35200585 probe_10 -0.49024387 probe_11 -1.09087811 probe_12 0.22008832 probe_13 2.54263342 probe_14 3.71106614 probe_15 2.12580554 probe_16 -0.06729251 pset at se.probe.coefs[[1]] Overall probe_1 0.06124122 probe_2 0.06039453 probe_3 0.06180433 probe_4 0.05948503 probe_5 0.06727454 probe_6 0.06016827 probe_7 0.06233682 probe_8 0.06791376 probe_9 0.05960599 probe_10 0.05963511 probe_11 0.05868359 probe_12 0.06046023 probe_13 0.05885199 probe_14 0.05829506 probe_15 0.05837877 probe_16 0.06340662 however, i'm unsure how to proceed from now on to decide whether a particular probeset is more "affected" by probe-specific effects than other probeset. any suggestion would be highly appreciated, thanks, robert.
probe affy affyPLM probe affy affyPLM • 1.3k views
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@matthew-mccall-4459
Last seen 5.5 years ago
United States
Robert, I'm not sure exactly what you're after, but you might want to look at the hgu133afrmavecs and hgu133plus2frmavecs data packages. The probe-effect (probeVec), within-batch residual variance (probeVarWithin), the between-batch residual variance (probeVarBetween), and the within probeset standard deviation (probesetSD) have all been computed using a large biologically diverse data set. Best, Matt On Mon, Mar 5, 2012 at 1:22 PM, Robert Castelo <robert.castelo at="" upf.edu=""> wrote: > dear list, > > i'm searching for a way to rank affy probesets from classical 3' affy > arrays by their probe effect magnitude. i mean that i would like to know > if a probeset is has a larger probe-specific effect than another one. > > i guess the solution should be in the affyPLM package since if i do > > library(affy) > library(affyPLM) > > ab <- ReadAffy() > pset <- fitPLM(ab) > > > i obtain an object (pset) of the PLMset class which contains slots > 'probe.coefs' and 'se.probe.coefs', where each is a list as many keys as > probesets and where each probeset contains information on the probe > effect of each probe within the probeset: > > head(names(pset at probe.coefs)) > [1] "1000_at" ? "1001_at" ? "1002_f_at" "1003_s_at" "1004_at" > "1005_at" > head(names(pset at se.probe.coefs)) > [1] "1000_at" ? "1001_at" ? "1002_f_at" "1003_s_at" "1004_at" > "1005_at" > > pset at probe.coefs[[1]] > ? ? ? ? ? ? Overall > probe_1 ? 0.97287528 > probe_2 ? 0.61454806 > probe_3 ?-2.81701693 > probe_4 1.68063395 > probe_5 ?-3.31991235 > probe_6 1.56657388 > probe_7 ?-3.30256264 > probe_8 -1.99431231 > probe_9 -0.35200585 > probe_10 -0.49024387 > probe_11 -1.09087811 > probe_12 0.22008832 > probe_13 2.54263342 > probe_14 ?3.71106614 > probe_15 ?2.12580554 > probe_16 -0.06729251 > > pset at se.probe.coefs[[1]] > ? ? ? ? ? ?Overall > probe_1 ?0.06124122 > probe_2 ?0.06039453 > probe_3 ?0.06180433 > probe_4 ?0.05948503 > probe_5 0.06727454 > probe_6 0.06016827 > probe_7 0.06233682 > probe_8 0.06791376 > probe_9 0.05960599 > probe_10 0.05963511 > probe_11 0.05868359 > probe_12 0.06046023 > probe_13 0.05885199 > probe_14 0.05829506 > probe_15 0.05837877 > probe_16 0.06340662 > > however, i'm unsure how to proceed from now on to decide whether a > particular probeset is more "affected" by probe-specific effects than > other probeset. any suggestion would be highly appreciated, > > thanks, > robert. > > _______________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor -- Matthew N McCall, PhD 112 Arvine Heights Rochester, NY 14611 Cell: 202-222-5880
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Matt, does probeVec represents some sort of baseline expression for each specific probe? does it make sense to calculate these vectors for another affy chip with 20 samples? thanks! robert. On 3/5/12 7:52 PM, Matthew McCall wrote: > Robert, > > I'm not sure exactly what you're after, but you might want to look at > the hgu133afrmavecs and hgu133plus2frmavecs data packages. The > probe-effect (probeVec), within-batch residual variance > (probeVarWithin), the between-batch residual variance > (probeVarBetween), and the within probeset standard deviation > (probesetSD) have all been computed using a large biologically diverse > data set. > > Best, > Matt > > On Mon, Mar 5, 2012 at 1:22 PM, Robert Castelo<robert.castelo at="" upf.edu=""> wrote: >> dear list, >> >> i'm searching for a way to rank affy probesets from classical 3' affy >> arrays by their probe effect magnitude. i mean that i would like to know >> if a probeset is has a larger probe-specific effect than another one. >> >> i guess the solution should be in the affyPLM package since if i do >> >> library(affy) >> library(affyPLM) >> >> ab<- ReadAffy() >> pset<- fitPLM(ab) >> >> >> i obtain an object (pset) of the PLMset class which contains slots >> 'probe.coefs' and 'se.probe.coefs', where each is a list as many keys as >> probesets and where each probeset contains information on the probe >> effect of each probe within the probeset: >> >> head(names(pset at probe.coefs)) >> [1] "1000_at" "1001_at" "1002_f_at" "1003_s_at" "1004_at" >> "1005_at" >> head(names(pset at se.probe.coefs)) >> [1] "1000_at" "1001_at" "1002_f_at" "1003_s_at" "1004_at" >> "1005_at" >> >> pset at probe.coefs[[1]] >> Overall >> probe_1 0.97287528 >> probe_2 0.61454806 >> probe_3 -2.81701693 >> probe_4 1.68063395 >> probe_5 -3.31991235 >> probe_6 1.56657388 >> probe_7 -3.30256264 >> probe_8 -1.99431231 >> probe_9 -0.35200585 >> probe_10 -0.49024387 >> probe_11 -1.09087811 >> probe_12 0.22008832 >> probe_13 2.54263342 >> probe_14 3.71106614 >> probe_15 2.12580554 >> probe_16 -0.06729251 >> >> pset at se.probe.coefs[[1]] >> Overall >> probe_1 0.06124122 >> probe_2 0.06039453 >> probe_3 0.06180433 >> probe_4 0.05948503 >> probe_5 0.06727454 >> probe_6 0.06016827 >> probe_7 0.06233682 >> probe_8 0.06791376 >> probe_9 0.05960599 >> probe_10 0.05963511 >> probe_11 0.05868359 >> probe_12 0.06046023 >> probe_13 0.05885199 >> probe_14 0.05829506 >> probe_15 0.05837877 >> probe_16 0.06340662 >> >> however, i'm unsure how to proceed from now on to decide whether a >> particular probeset is more "affected" by probe-specific effects than >> other probeset. any suggestion would be highly appreciated, >> >> thanks, >> robert. >> >> _______________________________________________ >> Bioconductor mailing list >> Bioconductor at r-project.org >> https://stat.ethz.ch/mailman/listinfo/bioconductor >> Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor > > >
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@james-w-macdonald-5106
Last seen 14 minutes ago
United States
Hi Robert, On 3/5/2012 1:22 PM, Robert Castelo wrote: > dear list, > > i'm searching for a way to rank affy probesets from classical 3' affy > arrays by their probe effect magnitude. i mean that i would like to know > if a probeset is has a larger probe-specific effect than another one. > > i guess the solution should be in the affyPLM package since if i do > > library(affy) > library(affyPLM) > > ab<- ReadAffy() > pset<- fitPLM(ab) > > > i obtain an object (pset) of the PLMset class which contains slots > 'probe.coefs' and 'se.probe.coefs', where each is a list as many keys as > probesets and where each probeset contains information on the probe > effect of each probe within the probeset: > > head(names(pset at probe.coefs)) > [1] "1000_at" "1001_at" "1002_f_at" "1003_s_at" "1004_at" > "1005_at" > head(names(pset at se.probe.coefs)) > [1] "1000_at" "1001_at" "1002_f_at" "1003_s_at" "1004_at" > "1005_at" > > pset at probe.coefs[[1]] > Overall > probe_1 0.97287528 > probe_2 0.61454806 > probe_3 -2.81701693 > probe_4 1.68063395 > probe_5 -3.31991235 > probe_6 1.56657388 > probe_7 -3.30256264 > probe_8 -1.99431231 > probe_9 -0.35200585 > probe_10 -0.49024387 > probe_11 -1.09087811 > probe_12 0.22008832 > probe_13 2.54263342 > probe_14 3.71106614 > probe_15 2.12580554 > probe_16 -0.06729251 > > pset at se.probe.coefs[[1]] > Overall > probe_1 0.06124122 > probe_2 0.06039453 > probe_3 0.06180433 > probe_4 0.05948503 > probe_5 0.06727454 > probe_6 0.06016827 > probe_7 0.06233682 > probe_8 0.06791376 > probe_9 0.05960599 > probe_10 0.05963511 > probe_11 0.05868359 > probe_12 0.06046023 > probe_13 0.05885199 > probe_14 0.05829506 > probe_15 0.05837877 > probe_16 0.06340662 > > however, i'm unsure how to proceed from now on to decide whether a > particular probeset is more "affected" by probe-specific effects than > other probeset. any suggestion would be highly appreciated, I'm not sure what you are looking to do with these data, but remember that the probe-specific effects in RMA are estimated as a nuisance variable, which are then excluded from computation of the expression value. So by definition the probesets should not be affected by probe-specific effects. Best, Jim > > thanks, > robert. > > _______________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor -- James W. MacDonald, M.S. Biostatistician University of Washington Environmental and Occupational Health Sciences 4225 Roosevelt Way NE, # 100 Seattle WA 98105-6099
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hi Jim, On 3/5/12 8:02 PM, James W. MacDonald wrote: [...] > I'm not sure what you are looking to do with these data, but remember > that the probe-specific effects in RMA are estimated as a nuisance > variable, which are then excluded from computation of the expression > value. So by definition the probesets should not be affected by > probe-specific effects. my understanding is that probesets in a microarray may have different baseline expression levels due to probe-specific effects. if i recall correctly this is illustrated in Fig. 1 of Zilliox and Irizarry (Nat. Meth., 2007) and this, for instance, complicates the question of determining whether a gene is expressed or not, which is tackled on that paper. i would like to assess somehow the agreement of these possibly different expression baselines between different genes and i thought that probe-specific effects would contain such information. to be more specific, if we would assume a simple additive model y_ij = probe_effect_i + sample_effect_j + noise_ij i'd like to know what genes have more similar or more disparate probe effects estimated as the probe_effect_i term in the previous linear model and i thought such information could be extracted from PLMset objects. cheers, robert.
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Robert, The probe-effects (probeVec) are not a baseline expression -- they are constrained to sum to zero within each probeset, so they represent the typical difference from the overall probeset expression. Matt On Mon, Mar 5, 2012 at 5:14 PM, Robert Castelo <robert.castelo at="" upf.edu=""> wrote: > hi Jim, > > On 3/5/12 8:02 PM, James W. MacDonald wrote: > [...] > > >> I'm not sure what you are looking to do with these data, but remember >> that the probe-specific effects in RMA are estimated as a nuisance >> variable, which are then excluded from computation of the expression >> value. So by definition the probesets should not be affected by >> probe-specific effects. > > > my understanding is that probesets in a microarray may have different > baseline expression levels due to probe-specific effects. if i recall > correctly this is illustrated in Fig. 1 of Zilliox and Irizarry (Nat. Meth., > 2007) and this, for instance, complicates the question of determining > whether a gene is expressed or not, which is tackled on that paper. > > i would like to assess somehow the agreement of these possibly different > expression baselines between different genes and i thought that > probe-specific effects would contain such information. > > to be more specific, if we would assume a simple additive model > > y_ij = probe_effect_i + sample_effect_j + noise_ij > > i'd like to know what genes have more similar or more disparate probe > effects estimated as the probe_effect_i term in the previous linear model > and i thought such information could be extracted from PLMset objects. > > cheers, > > robert. > > _______________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: > http://news.gmane.org/gmane.science.biology.informatics.conductor -- Matthew N McCall, PhD 112 Arvine Heights Rochester, NY 14611 Cell: 202-222-5880
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