Statistical analysis of real-time PCR data

BMC Bioinformatics
Joshua S YuanC Neal Stewart

Abstract

Even though real-time PCR has been broadly applied in biomedical sciences, data processing procedures for the analysis of quantitative real-time PCR are still lacking; specifically in the realm of appropriate statistical treatment. Confidence interval and statistical significance considerations are not explicit in many of the current data analysis approaches. Based on the standard curve method and other useful data analysis methods, we present and compare four statistical approaches and models for the analysis of real-time PCR data. In the first approach, a multiple regression analysis model was developed to derive DeltaDeltaCt from estimation of interaction of gene and treatment effects. In the second approach, an ANCOVA (analysis of covariance) model was proposed, and the DeltaDeltaCt can be derived from analysis of effects of variables. The other two models involve calculation DeltaCt followed by a two group t-test and non-parametric analogous Wilcoxon test. SAS programs were developed for all four models and data output for analysis of a sample set are presented. In addition, a data quality control model was developed and implemented using SAS. Practical statistical solutions with SAS programs were developed for real-time P...Continue Reading

References

Oct 1, 1996·Genome Research·C A HeidP M Williams
Oct 1, 1996·Genome Research·U E GibsonP M Williams
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
Jun 18, 2002·Trends in Molecular Medicine·Dieter Klein
Oct 26, 2002·BMC Biotechnology·Giovanna MasonGian Paolo Accotto
Jan 10, 2003·Nucleic Acids Research·Filip PattynJo Vandesompele
May 24, 2003·Trends in Molecular Medicine·Simone MocellinFrancesco M Marincola
Jul 11, 2003·Nucleic Acids Research·Stuart N PeirsonRussell G Foster
Oct 8, 2003·Nucleic Acids Research·Ales TichopadMichael W Pfaffl
Dec 16, 2004·BioTechniques·Peyton CookKenton S Miller
Jun 16, 2005·Journal of Molecular Endocrinology·S A BustinM W Pfaffl
Sep 17, 2005·Archives of Biochemistry and Biophysics·Yue YangFeng Chen

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Citations

Oct 15, 2008·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Dilip R PantheeC Neal Stewart
Jan 21, 2010·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Doug J HinchliffeBarbara A Triplett
Nov 20, 2009·Current Microbiology·Nicholas C ButzinMary Lynne Perille Collins
Jul 27, 2007·Breast Cancer Research and Treatment·Nalin SiriwardhanaHwa-Chain Robert Wang
Apr 3, 2010·Journal of Molecular Histology·Augusto B ReisFernando M Reis
Dec 15, 2011·Pharmaceutical Research·Christopher C CossJames T Dalton
Jan 2, 2009·In Vitro Cellular & Developmental Biology. Animal·Shaneen S BaxterMichael J Fay
Apr 28, 2011·In Vitro Cellular & Developmental Biology. Animal·Zohreh MakoolatiMehdi Forouzandeh-Moghadam
May 28, 2010·Journal of Molecular Neuroscience : MN·Gabriela BedóAna Aranda
May 31, 2013·Clinical & Translational Oncology : Official Publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico·Y KılıçM Sakızlı
Sep 4, 2012·Journal of Physiology and Biochemistry·Christopher R LindholmKurt W Saupe
Apr 16, 2013·Comparative Immunology, Microbiology and Infectious Diseases·Amaia LarruskainBegoña M Jugo
Nov 12, 2013·Protein Expression and Purification·Sebastian A StifterNicole A de Weerd
Feb 1, 2010·Animal : an International Journal of Animal Bioscience·A CánovasR N Pena
Jun 23, 2011·Chemical Research in Toxicology·Giuseppina AmodioNicola Zambrano
May 13, 2011·Gene Therapy·B M HoveyD C Seldin
Aug 12, 2009·Nature·Han LiManuel Serrano
Jul 23, 2013·Nature Immunology·Nicole A de WeerdPaul J Hertzog
Sep 7, 2012·Reproduction, Fertility, and Development·Maryam PashaiaslPaul J Verma
Jun 14, 2012·Proceedings of the National Academy of Sciences of the United States of America·Agenor LimonRicardo Miledi
Jan 2, 2013·Proceedings of the National Academy of Sciences of the United States of America·Dominique PromeneurPeter Agre
May 4, 2011·The Journal of Biological Chemistry·Jeremie Vidal-DupiolGuillaume Mitta
Nov 30, 2011·The Journal of Biological Chemistry·Hao WangGuoqing Sheng
Oct 11, 2012·Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment·Ali A AlshatwiAbdulrahman A Alwarthan
Jul 12, 2013·Journal of Computational Biology : a Journal of Computational Molecular Cell Biology·Xiayu RaoXuelin Huang
Jan 22, 2011·Nucleic Acids Research·Narendra Pratap SinghRakesh K Mishra
Jul 5, 2013·Plant & Cell Physiology·Jong-Pil HongRyoung Shin

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Methods Mentioned

BETA
PCR
transgenic
PCRs

Software Mentioned

SAS Macro
Program4
MathType
Excel
QC
MR
ANCOVA
SAS
SASOutputs
Program2

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