Qpcr And Error Bar
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fold-change data? I'm considering some real time data on tissues treated and untreated upon a given stress. Obviously based on three technical replicate in each condition
Fold Change Error Bars
I obtained a normalized expression value associated with a standard error based on standard deviation of fold change the replicates. If I calculate the log2 of the fold change (treated / untreated) how can I calculate
Qpcr Fold Change Standard Deviation
the corresponding error bars? Topics Basic Statistical Methods × 401 Questions 93 Followers Follow Real-Time PCR × 2,152 Questions 3,371 Followers Follow Transcriptomics × 397 Questions 17,825 Followers Follow Gene Expression qpcr biological replicates standard deviation × 1,722 Questions 25,297 Followers Follow Apr 17, 2014 Share Facebook Twitter LinkedIn Google+ 0 / 0 Popular Answers Jochen Wilhelm · Justus-Liebig-Universität Gießen I suppose you have a standard error of the mean log expression for each condition se_A and se_B. The log ratio is the difference between the log expressions, and the standard error of this difference is given qpcr data analysis error bars by sqrt(se_A²+se_B²)*. *assuming A and B are uncorrelated See: http://en.wikipedia.org/wiki/Propagation_of_uncertainty Apr 17, 2014 All Answers (7) Jochen Wilhelm · Justus-Liebig-Universität Gießen I suppose you have a standard error of the mean log expression for each condition se_A and se_B. The log ratio is the difference between the log expressions, and the standard error of this difference is given by sqrt(se_A²+se_B²)*. *assuming A and B are uncorrelated See: http://en.wikipedia.org/wiki/Propagation_of_uncertainty Apr 17, 2014 Jo Vandesompele · Ghent University All formulas for error propagation during qPCR data-analysis are mentioned in attached paper. The formulas are integrated in Biogazelle's qbase+ software (http://www.qbaseplus.com). Source Available from: Jo Vandesompele Article: Hellemans J, Mortier GR, De Paepe A, Speleman F, Vandesompele JqBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol 8:R19 Jan Hellemans · Geert Mortier · Anne De Paepe · [...] · Jo Vandesompele [Show abstract] [Hide abstract] ABSTRACT: Although quantitative PCR (qPCR) is becoming the method of choice for expression profiling of selected genes, accurate and straightforward processing of the raw measurements remains a major hurdle. Here we out
a bar graph? I have my raw data according to two groups of different types of animals that are being tested with pharmacological compounds or PBS. I am using
Standard Error Qpcr
the 2deltaCT method. I've seen different papers with many ways of graphing results, how to plot qpcr data lets say for instance: relative expression vs gene of interest, fold change vs gene of interest, RQ vs Gene. Does anyone
How To Present Qpcr Data
have an opinion on the best way to depict your relative quantification data? Topics Molecular Biological Techniques × 7,058 Questions 33,958 Followers Follow PCR × 5,022 Questions 71,924 Followers Follow Methods × 3,976 https://www.researchgate.net/post/Can_anyone_help_with_calculating_error_in_RT-qPCRs_fold-change_data Questions 132,080 Followers Follow Gene Expression × 1,722 Questions 25,297 Followers Follow Real-Time PCR × 2,152 Questions 3,371 Followers Follow Feb 27, 2014 Share Facebook Twitter LinkedIn Google+ 0 / 1 Popular Answers Jochen Wilhelm · Justus-Liebig-Universität Gießen Short answer: Corectly applied, barcharts only allow you to show ddCt values. Longer: If you want to relate expressions among several groups, it's best to show dCt. If the https://www.researchgate.net/post/How_do_I_publish_qPCR_data_in_a_bar_graph comparisons at all done to the same reference, best show ddCt. Never ever use simple barcharts (as used so terribly often in biomedical papers). If you have small sample sizes, you can show dCt values simply in 1D scatterplots. For larger sample sizes, dCt values can be shown in boxplots, or in dot-plots with error bars (indicating median and IQR or the mean and 95%CI). In case of ddCt values the only option are dot-plots with mean and CI (since there are no "individual measurements" and you can only provide the mean ddCt). If you are forced to show 2^dCt or 2^ddCt, then I'd suggest to calculate all statistics (like medians, IQRs, means, CIs) for the dCt (or ddCt) values and potentiate these results to show them in a plot. For the means and CIs this gives you the "geometric means" with according CI (what is not symmetric around the mean). If you are forced to use barcharts, you cannot show dCt because this quantity has no interpretable zero value (so a hight of a bar, even the direction [positive or negative] provides no interpretable information; so the purpose of the barchart [=showing bar areas] is completely off-topic). You
biological replicate - (Dec/20/2011 )Visit this topic http://www.protocol-online.org/biology-forums-2/posts/23815.html in live forum Printer Friendly VersionABI has a step by step guide for qPCR statistics that ultimately gives you the fold change and the standard deviation (can be found here). My assumption is that the calculated SD only represents the variation in error bar technical replicates and not the biological replicates. How would you incorporate the variations in both technical and biological replicates into the final error bars (say SD)? Is the technical replicate SD is reflected in the final result at all? any input is qpcr and error highly appreciated. Kaveh -kaveh- From what I have seen, most people just plot technical replicates +/- error bars (95% CIs) and then they indicate that they repeated the experiment 2 or 3 times and had similar results. I think for biological replicates you are correct in assuming that you just ignore the SD for the technical replicates and just calculate the mean and SD for the biological replicates. -doxorubicin- Yes, we take SD of technical replicates as a indication how precise was the experiment, but if we plott biological replicates, we only calculate SD for the biological ones and ignore the technical. -Trof- Visit this topic in BioForum Printer Friendly Version About Terms of Service Privacy Feedback Sponsorship © 1999-2013 Protocol Online, All rights reserved.