HTqPCR
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Heidi Dvinge ★ 2.0k
@heidi-dvinge-2195
Last seen 9.6 years ago
Hello Arun, > Dear Heidi > Iam using HTqPCR for my 384 well TLDA data analysis. Iam analysis 40 > patient sample and 20 normal samples. For my analysis is it possible > to get the fold change of that 40 patient samples individually > campared to mean of all normal. at present analysis iam getting mean > fold change of all patient compared to all normal. > if i can get that i can find which all patients have higher value than > normal. > There's no default way of doing this in HTqPCR, but you can easily do it 'manually' by extracting the Ct values. If you haven't already done it, you should consider removing unreliable Ct values first, to avoid them biasing the results. For example: # Load example data data(qPCRpros) # Mark unreliable values, if this isn't already done q.tmp <- setCategory(qPCRpros, Ct.max = 35, Ct.min = 10, quantile=NULL) q.filter <- filterCategory(q.tmp) # Count number of NAs, just FYI colSumsis.na(getCt(q.filter))) # In this example, I assume that the first 3 samples are the controls control.index <- sampleNames(q.filter) %in% c("sample1", "sample2", "sample3") # Get the mean of these tmp <- getCt(q.filter)[,control.index] control.mean <- rowMeans(tmp, na.rm=TRUE) # Comapre to all the non-controls tmp <- getCt(q.filter)[,!control.index] sample.vs.control <- tmp-control.mean > secondly is it possible to get the excell table data of normalized ct > values. > Yep, you can export these using the standard write.table() function: # Get Ct values tmp <- getCt(qPCRpros) # Write to file write.table(tmp, file="normalized_Ct_values.xls", sep="\t") HTH \Heidi > Deepak > + 91 471 2522338 >
HTqPCR HTqPCR • 788 views
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