Hi Jo I am doing expression profile and when I am making my standard curve I am using 1x, 1/2, 1/4, 1/8, 1/16 dilution series since my cDNA amount is limiting....
Hi, I have been running real time reactions on ABI-7300. For one particular sample I had 2 replicates and ran both the replicates in duplicates. But both the...
Hi All, Just wondering is there a difference between the way GeNorm caluclates M value in comparison to Qbase. When I calculate with GeNorm I get M values of...
Dear Stephen Both qBase and geNorm use the same formula to calculate the M value. You do realize that the input is different for geNorm (should be relative ...
please note that we are talking about relative quantification, so the units are arbitrary units or 'fold change compared to the calibrator sample', or if you...
Dear Jo, Yes, my input for geNorm was the relative quantities, I divided the lowest CT value by every other sample. I have also double checked that its the...
Dear Stephen I'm afraid your calculations are not correct. You are not supposed to divide Ct values. Please take a look at the example calculation file on the...
Dear Jo, Thanks for your help, for some reason I was using the ratios of lowest CT to other sample CT instead of relative quantities. Cheers, Stephen ... to ...
Hi, Just a quick question for the group. What are the main differences between version 1.3.3 and 1.3.4? I'm particularly interested in the computation...
Am very impressed with qBase analysis of data file examples, but have had no success trying to format data from an Mx4000.From the manual it is clear that...
Hi! I've been trying to use qBase for the last couple weeks. After getting all my input files correctly formatted and figuring out how to use the program I am...
qBase is a relative quantification program employing universally applicable and advanced quantification models. Rescaling is performed on a gene per gene...
Hi! I saw your webpage and thought that qBase may provide tools that I need. I have a set of customized (sort of) qPCR data, which essentially is the...
Any table of Ct values that can be converted to any of the supported import types (see descriptions at the end of the manual) can be used by qBase after...
Hi Jan, Thank you for the prompt reply! Actually my data consist of the raw intensity with cycle numbers and need to calculate Ct values for each condition....
We released a new version of qBase (minor bug fixes), along with a step-by-step tutorial detailing the different steps in the workflow (this is different than...
"qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data" by Hellemans et al. is...
Dear Jo I am begining a fairly large qPCR study, but I am concerned that we may not be able to be able to analyse or data to it's full potential if we get the...
... Of course! The sample maximization approach is only proposed to reduced inter-run variation when comparing samples. You are still able to compare genes...
Thank you for your reply Jo You mention the use of IRCs in your reply. Can you clarify some points regarding IRCs for me please. In the qBase tutorial, the...
... Indeed, because all samples fit on the same plate. ... Correct. ... There is no run-to-run variatio, because all samples are tested in the same plate....
Greetings, My last question was stupid. I have used Opticon Monitor to output Ct values for me, inserted these values into QBase format files, and then used...
Hello Im very new to the field of qRT-PCR analylis and would like to use qBase for my calculations. But i encountered a, lets call it "data organisation...
Can you also state the number of samples in each subgroup and number of PCR replicates? What are your inter-run calibrators? How many IRCs do you have? If...
Hello Jo Thank you for your quick response. I have 8 samples in each of the 4 subgoups. All the samples ran as duplicates. KO-plate setup: - triplicate...
Your experimental design seems perfect. I just wanted to note that you can also use the standards for the IRC procedure. If these are diluted cDNAs (pooled or...
QBase detects IRCs that are really not IRCs. They are normal samples. It only detects these false IRCs in the last Run I enter (I am entering 5 runs into my...
I have been able to solve the run time error "9" that occured while detecting IRCs after files had been imported. The solution was to make all gene and sample...
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