Journal ArticleDOI
Analyzing real-time PCR data by the comparative C(T) method.
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TLDR
This protocol provides an overview of the comparative CT method for quantitative gene expression studies and various examples to present quantitative gene Expression data using this method.Abstract:
Two different methods of presenting quantitative gene expression exist: absolute and relative quantification. Absolute quantification calculates the copy number of the gene usually by relating the PCR signal to a standard curve. Relative gene expression presents the data of the gene of interest relative to some calibrator or internal control gene. A widely used method to present relative gene expression is the comparative C(T) method also referred to as the 2 (-DeltaDeltaC(T)) method. This protocol provides an overview of the comparative C(T) method for quantitative gene expression studies. Also presented here are various examples to present quantitative gene expression data using this method.read more
Citations
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A major role of insulin in promoting obesity- associated adipose tissue inflammation
David J. Pedersen,Adilson L. Guilherme,Laura V. Danai,Lauren Heyda,Anouch Matevossian,Jessica Cohen,Sarah M. Nicoloro,Juerg R. Straubhaar,Hye Lim Noh,Dae Young Jung,Jason K. Kim,Michael P. Czech +11 more
TL;DR: It is indicated that obesity-associated hyperinsulinemia unexpectedly drives AT inflammation in obese mice, which in turn contributes to factors that suppress insulin-stimulated adipocyte DNL and systemic insulin sensitivity.
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Poor Prognosis Indicated by Venous Circulating Tumor Cell Clusters in Early-Stage Lung Cancers.
Vasudha Murlidhar,Rishindra M. Reddy,Shamileh Fouladdel,Lili Zhao,Martin K. Ishikawa,Svetlana Grabauskiene,Zhuo Zhang,Jules Lin,Andrew C. Chang,Philip W. Carrott,William R. Lynch,Mark B. Orringer,Chandan Kumar-Sinha,Nallasivam Palanisamy,David G. Beer,Max S. Wicha,Nithya Ramnath,Ebrahim Azizi,Sunitha Nagrath +18 more
TL;DR: CTCs isolated from early stages of lung cancer are predictive of poor prognosis and can be interrogated to determine biomarkers predictive of recurrence, suggesting survival advantage of clusters in circulation.
Journal ArticleDOI
Dissecting the mechanisms responsible for the multiple insecticide resistance phenotype in Anopheles gambiae s.s., M form, from Vallée du Kou, Burkina Faso
Rachel M. Kwiatkowska,Naomi Platt,Rodolphe Poupardin,Helen Irving,Roch K. Dabiré,Sara N. Mitchell,Christopher M. Jones,Abdoulaye Diabaté,Hilary Ranson,Charles S. Wondji +9 more
TL;DR: High prevalence of resistance in Vallée du Kou to pyrethroids, DDT and dieldrin, moderate level for carbamates and full susceptibility to organophosphates is detected, highlighting the complex network of mechanisms conferring multiple resistance in malaria vectors.
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Epigenetically-driven anatomical diversity of synovial fibroblasts guides joint-specific fibroblast functions
Mojca Frank-Bertoncelj,Michelle Trenkmann,Katja Klein,Emmanuel Karouzakis,Hubert Rehrauer,A. Bratus,Christoph Kolling,Maria Armaka,Andrew Filer,Beat A. Michel,Christopher D. Buckley,George Kollias,Caroline Ospelt +12 more
TL;DR: It is shown that transcriptomic differences in synovial fibroblasts from different joint locations are shown and that HOX gene signatures reflect the joint-specific origins of mouse and human synovials, indicating that local stroma might control positional disease patterns in arthritis but in any disease with a prominent stromal component.
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β-Hydroxybutyrate Modulates N-Type Calcium Channels in Rat Sympathetic Neurons by Acting as an Agonist for the G-Protein-Coupled Receptor FFA3
TL;DR: The results indicate that endogenous BHB levels, especially when elevated under certain conditions, such as starvation, diabetic ketoacidosis, and ketogenic diets, play a potentially important role in regulating the activity of the SNS through FFA3.
References
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Journal ArticleDOI
Analysis of relative gene expression data using real-time quantitative pcr and the 2(-delta delta c(t)) method
TL;DR: The 2-Delta Delta C(T) method as mentioned in this paper was proposed to analyze the relative changes in gene expression from real-time quantitative PCR experiments, and it has been shown to be useful in the analysis of realtime, quantitative PCR data.
Journal ArticleDOI
A new mathematical model for relative quantification in real-time RT-PCR.
TL;DR: This study enters into the particular topics of the relative quantification in real-time RT-PCR of a target gene transcript in comparison to a reference gene transcript and presents a new mathematical model that needs no calibration curve.
Journal ArticleDOI
Cluster analysis and display of genome-wide expression patterns
TL;DR: A system of cluster analysis for genome-wide expression data from DNA microarray hybridization is described that uses standard statistical algorithms to arrange genes according to similarity in pattern of gene expression, finding in the budding yeast Saccharomyces cerevisiae that clustering gene expression data groups together efficiently genes of known similar function.
Journal ArticleDOI
A Multigene Assay to Predict Recurrence of Tamoxifen-Treated, Node-Negative Breast Cancer
Soonmyung Paik,Steven Shak,Gong Tang,Chungyeul Kim,Joffre B. Baker,Maureen T. Cronin,Frederick L. Baehner,Michael G. Walker,Drew Watson,Taesung Park,William Hiller,Edwin R. Fisher,D. Lawrence Wickerham,John Bryant,Norman Wolmark +14 more
TL;DR: The recurrence score has been validated as quantifying the likelihood of distant recurrence in tamoxifen-treated patients with node-negative, estrogen-receptor-positive breast cancer and could be used as a continuous function to predict distant recurrent in individual patients.
Journal ArticleDOI
Real-time quantification of microRNAs by stem–loop RT–PCR
Caifu Chen,Dana Ridzon,Adam Broomer,Zhaohui Zhou,Danny H. Lee,Julie T. Nguyen,Maura Barbisin,Nan Lan Xu,Vikram R. Mahuvakar,Mark R. Andersen,Kaiqin Lao,Kenneth J. Livak,Karl J. Guegler +12 more
TL;DR: A novel microRNA quantification method has been developed using stem–loop RT followed by TaqMan PCR analysis, which enables fast, accurate and sensitive miRNA expression profiling and can identify and monitor potential biomarkers specific to tissues or diseases.