problem with qPCR raw data
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@alberto-goldoni-3412
Last seen 9.6 years ago
Dear all, there is a method to calculate the Ct value if i have the file.txt with the fuolescence intensity like this one: Raw Data Experiment - PIASTRA 1 ALL+CML (Run on LCS480 1.2.9.11) SamplePos SampleName Prog# Seg# Cycle# Time Temp 483-533 A1 4200_ABL 2 2 1 420616 59.93 6.67 A1 4200_ABL 2 2 2 529833 59.93 6.72 A1 4200_ABL 2 2 3 639216 59.96 6.86 A1 4200_ABL 2 2 4 748850 59.96 6.94 A1 4200_ABL 2 2 5 858416 59.96 7.53 A1 4200_ABL 2 2 6 967600 59.96 7.59 A1 4200_ABL 2 2 7 1076816 59.96 7.64 A1 4200_ABL 2 2 8 1186033 59.93 7.69 A1 4200_ABL 2 2 9 1295616 59.96 7.74 A1 4200_ABL 2 2 10 1404816 59.96 7.77 A1 4200_ABL 2 2 11 1513816 59.96 7.79 A1 4200_ABL 2 2 12 1623000 59.96 7.86 A1 4200_ABL 2 2 13 1732650 59.96 7.87 A1 4200_ABL 2 2 14 1841800 59.96 7.92 A1 4200_ABL 2 2 15 1951033 59.96 7.93 A1 4200_ABL 2 2 16 2060000 59.96 7.95 A1 4200_ABL 2 2 17 2169216 59.96 7.99 A1 4200_ABL 2 2 18 2278233 59.96 8 A1 4200_ABL 2 2 19 2387216 59.93 8 A1 4200_ABL 2 2 20 2496433 59.96 8.03 A1 4200_ABL 2 2 21 2605616 59.93 8.05 A1 4200_ABL 2 2 22 2714816 59.96 8.06 A1 4200_ABL 2 2 23 2824016 59.96 8.08 A1 4200_ABL 2 2 24 2933183 59.96 8.12 A1 4200_ABL 2 2 25 3042433 59.96 8.16 A1 4200_ABL 2 2 26 3151633 59.93 8.22 A1 4200_ABL 2 2 27 3260833 59.96 8.3 A1 4200_ABL 2 2 28 3370016 59.93 8.42 -- ---------------------------------------- Alberto Goldoni, PhD IT Lan Manager Medical Genetics Unit S. Orsola-Malpighi Hospital Via Massarenti n.9, Pad 11 40100 Bologna, Italy ---------------------------------------- -------------- next part -------------- An embedded and charset-unspecified text was scrubbed... Name: PIASTRA 1 ALL+CML.txt URL: <https: stat.ethz.ch="" pipermail="" bioconductor="" attachments="" 20100119="" 61f36417="" attachment.txt="">
Genetics Genetics • 815 views
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Heidi Dvinge ★ 2.0k
@heidi-dvinge-2195
Last seen 9.6 years ago
Hello Alberto, it's not in BioConductor, and I've never used it myself, but the package qpcR might be able to do what you're after. As far as I remember it's on CRAN, but if not you can get it from http://www.dr- spiess.de/qpcR.html Cheers \Heidi On 19 Jan 2010, at 11:09, Alberto Goldoni wrote: > Dear all, > there is a method to calculate the Ct value if i have the file.txt > with the fuolescence intensity like this one: > > Raw Data > Experiment - PIASTRA 1 ALL+CML (Run on LCS480 1.2.9.11) > > > > > SamplePos SampleName Prog# Seg# Cycle# Time Temp 483-533 > A1 4200_ABL 2 2 1 420616 59.93 6.67 > A1 4200_ABL 2 2 2 529833 59.93 6.72 > A1 4200_ABL 2 2 3 639216 59.96 6.86 > A1 4200_ABL 2 2 4 748850 59.96 6.94 > A1 4200_ABL 2 2 5 858416 59.96 7.53 > A1 4200_ABL 2 2 6 967600 59.96 7.59 > A1 4200_ABL 2 2 7 1076816 59.96 7.64 > A1 4200_ABL 2 2 8 1186033 59.93 7.69 > A1 4200_ABL 2 2 9 1295616 59.96 7.74 > A1 4200_ABL 2 2 10 1404816 59.96 7.77 > A1 4200_ABL 2 2 11 1513816 59.96 7.79 > A1 4200_ABL 2 2 12 1623000 59.96 7.86 > A1 4200_ABL 2 2 13 1732650 59.96 7.87 > A1 4200_ABL 2 2 14 1841800 59.96 7.92 > A1 4200_ABL 2 2 15 1951033 59.96 7.93 > A1 4200_ABL 2 2 16 2060000 59.96 7.95 > A1 4200_ABL 2 2 17 2169216 59.96 7.99 > A1 4200_ABL 2 2 18 2278233 59.96 8 > A1 4200_ABL 2 2 19 2387216 59.93 8 > A1 4200_ABL 2 2 20 2496433 59.96 8.03 > A1 4200_ABL 2 2 21 2605616 59.93 8.05 > A1 4200_ABL 2 2 22 2714816 59.96 8.06 > A1 4200_ABL 2 2 23 2824016 59.96 8.08 > A1 4200_ABL 2 2 24 2933183 59.96 8.12 > A1 4200_ABL 2 2 25 3042433 59.96 8.16 > A1 4200_ABL 2 2 26 3151633 59.93 8.22 > A1 4200_ABL 2 2 27 3260833 59.96 8.3 > A1 4200_ABL 2 2 28 3370016 59.93 8.42 > > > -- > ---------------------------------------- > Alberto Goldoni, PhD > IT Lan Manager > Medical Genetics Unit > S. Orsola-Malpighi Hospital > Via Massarenti n.9, Pad 11 > 40100 Bologna, Italy > ---------------------------------------- > > <piastra 1="" all+cml.txt="">_______________________________________________ > 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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Heidi Dvinge ★ 2.0k
@heidi-dvinge-2195
Last seen 9.6 years ago
Hi Andreia, the Ct values for qPCRset objects can be subset like any standard R- object, i.e. using the [,] notation. samplesNames() just gives you the names of the individual samples. If you want to plot e.g. sample 1 you can say: plot(exprsraw.cat)[,1], exprs(q.norm)[,1]) If you have for example 4 samples, you can plot them next to each other like in the vignette by changing some parameters in par(). par(mfrow=c(2,2), mar=c(2,2,2,1)) for (i in 1:4) { plot(exprsraw.cat)[,i], exprs(q.norm)[,i]) } If you mean with a different colour for each sample like in the vignette, then you can define a colour vector such as my.col <- rep(brewer.pal(n.samplesraw.cat), "Spectral"), each=n.wells raw.cat)) plot(exprsraw.cat), exprs(q.norm), pch=20, col=my.col) In the latest version of HTqPCR (1.1.2) the vignette includes a bit more about the raw versus normalised plots in figure 6, since I've had other requests for that. HTH \Heidi On 20 Jan 2010, at 11:37, Andreia Fonseca wrote: > Hi Heidi, > > sorry but I am back at HTqPCR again and I am trying to produce the > plots raw Ct versus normalized CT values. I am using the command > plot(exprsraw.cat)~exprs(q.norm)) > > but I would like to plot it per sample, like in the manual, I saw > that sampleNames() function but I am not managing to understand how > to put it in the plot command. Can you tell me how you did it? > > Cheers > Andreia > > On Tue, Jan 19, 2010 at 5:54 PM, Heidi Dvinge <heidi@ebi.ac.uk> wrote: > Hello Alberto, > > it's not in BioConductor, and I've never used it myself, but the > package qpcR might be able to do what you're after. As far as I > remember it's on CRAN, but if not you can get it from http://www.dr- > spiess.de/qpcR.html > > Cheers > \Heidi > > > > On 19 Jan 2010, at 11:09, Alberto Goldoni wrote: > > Dear all, > there is a method to calculate the Ct value if i have the file.txt > with the fuolescence intensity like this one: > > Raw Data > Experiment - PIASTRA 1 ALL+CML (Run on LCS480 1.2.9.11) > > > > > SamplePos SampleName Prog# Seg# Cycle# Time > Temp 483-533 > A1 4200_ABL 2 2 1 420616 59.93 6.67 > A1 4200_ABL 2 2 2 529833 59.93 6.72 > A1 4200_ABL 2 2 3 639216 59.96 6.86 > A1 4200_ABL 2 2 4 748850 59.96 6.94 > A1 4200_ABL 2 2 5 858416 59.96 7.53 > A1 4200_ABL 2 2 6 967600 59.96 7.59 > A1 4200_ABL 2 2 7 1076816 > 59.96 7.64 > A1 4200_ABL 2 2 8 1186033 > 59.93 7.69 > A1 4200_ABL 2 2 9 1295616 > 59.96 7.74 > A1 4200_ABL 2 2 10 1404816 > 59.96 7.77 > A1 4200_ABL 2 2 11 1513816 > 59.96 7.79 > A1 4200_ABL 2 2 12 1623000 > 59.96 7.86 > A1 4200_ABL 2 2 13 1732650 > 59.96 7.87 > A1 4200_ABL 2 2 14 1841800 > 59.96 7.92 > A1 4200_ABL 2 2 15 1951033 > 59.96 7.93 > A1 4200_ABL 2 2 16 2060000 > 59.96 7.95 > A1 4200_ABL 2 2 17 2169216 > 59.96 7.99 > A1 4200_ABL 2 2 18 2278233 > 59.96 8 > A1 4200_ABL 2 2 19 2387216 > 59.93 8 > A1 4200_ABL 2 2 20 2496433 > 59.96 8.03 > A1 4200_ABL 2 2 21 2605616 > 59.93 8.05 > A1 4200_ABL 2 2 22 2714816 > 59.96 8.06 > A1 4200_ABL 2 2 23 2824016 > 59.96 8.08 > A1 4200_ABL 2 2 24 2933183 > 59.96 8.12 > A1 4200_ABL 2 2 25 3042433 > 59.96 8.16 > A1 4200_ABL 2 2 26 3151633 > 59.93 8.22 > A1 4200_ABL 2 2 27 3260833 > 59.96 8.3 > A1 4200_ABL 2 2 28 3370016 > 59.93 8.42 > > > -- > ---------------------------------------- > Alberto Goldoni, PhD > IT Lan Manager > Medical Genetics Unit > S. Orsola-Malpighi Hospital > Via Massarenti n.9, Pad 11 > 40100 Bologna, Italy > ---------------------------------------- > > <piastra 1="" all+cml.txt="">_______________________________________________ > > Bioconductor mailing list > Bioconductor@stat.math.ethz.ch > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/ > gmane.science.biology.informatics.conductor > > _______________________________________________ > Bioconductor mailing list > Bioconductor@stat.math.ethz.ch > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/ > gmane.science.biology.informatics.conductor > > > > -- > -------------------------------------------- > Andreia J. Amaral > Unidade de Imunologia ClĂ­nica > Instituto de Medicina Molecular > Universidade de Lisboa > email: andreiaamaral@fm.ul.pt > andreia.fonseca@gmail.com [[alternative HTML version deleted]]
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