HTqPCR
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marco fabbri ▴ 320
@marco-fabbri-1657
Last seen 8.2 years ago
Italy
Dear all, I am running HTqPCR to analyse tilda. I run 3 TLDA, in each cards thre are 2 samples, but the samples are diffrerents in the cards card1: Control and LPS card2: LPS and PMA card3: Control and LPS If the three carder would have the same sample, I have no problem as shown in the vignette sample2a.order <- factor(rep(c("Control","LPS"), each=192)) qPCRnew2[1] <- changeCtLayout(raw, sample.order = sample2a.order) but my situation is different Can you help me? thank you marco
HTqPCR HTqPCR • 1.0k views
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Heidi Dvinge ★ 2.0k
@heidi-dvinge-2195
Last seen 10.3 years ago
Hello Marco, > Dear all, > > I am running HTqPCR to analyse tilda. > > I run 3 TLDA, in each cards thre are 2 samples, but the samples are > diffrerents in the cards > card1: Control and LPS > card2: LPS and PMA > card3: Control and LPS > Actually, it doesn't matter whether the samples are the same or not. Instead of having a data matrix with 384 x 3 values you want 192 x 6. The content of these 6 columns can vary. > > If the three carder would have the same sample, I have no problem as > shown in the vignette > sample2a.order <- factor(rep(c("Control","LPS"), each=192)) > qPCRnew2[1] <- changeCtLayout(raw, sample.order = sample2a.order) > The sample2a.order object doesn't have to contain the real sample names at all. It's merely to indicate where on the qPCR card the different samples are located. For example, you can imagine a card with 8 rows of wells. The two samples can then either be loaded into rows 1-4 and 5-8, or 1,3,5,7 and 2,4,6,8 depending on the layut of the card. In you case, you can therefore just rename the samples as required. E.g. data(qPCRraw) sample2a.order <- factor(rep(c("X","Y"), each=192)) qPCRnew <- changeCtLayout(qPCRraw[,1:3], sample.order = sample2a.order) sampleNames(qPCRnew) <- c("Control", "LPS", "LPS", "PMA", "Control", "LPS") In this case you'd probably want to use e.g. clusterCt(qPCRnew, type = "samples"), to check whether the re-formatted qPCRset object cluster by biological samples or by the original 3 cards. HTH \Heidi > but my situation is different > Can you help me? > > > thank you > > marco >
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