Reference Gene Selection and Evaluation for Gene Expression Studies Using qRT-PCR in the White-Backed Planthopper,Sogatella furcifera(Hemiptera: Delphacidae)

2015 ◽  
Vol 109 (2) ◽  
pp. 879-886 ◽  
Author(s):  
Xing-kui An ◽  
Mao-lin Hou ◽  
Yu-di Liu
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Gene Selection ◽  
Sogatella Furcifera ◽  
Reference Gene Selection ◽  
Qrt Pcr ◽  
Expression Studies ◽  
Gene Expression Studies

scholarly journals Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers

2011 ◽  
Vol 8 (1) ◽  
pp. 308 ◽  
Author(s):  
Guillermo A Maroniche ◽  
Mónica Sagadín ◽  
Vanesa C Mongelli ◽  
Graciela A Truol ◽  
Mariana del Vas
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Gene Selection ◽  
Reference Gene Selection ◽  
Expression Studies ◽  
Selection For ◽  
Gene Expression Studies

Impact of reference gene selection for type 2 cannabinoid receptor gene expression studies in human spermatozoa

Andrologia ◽  
2012 ◽  
Vol 45 (4) ◽  
pp. 278-284 ◽  
Author(s):  
A. A. Amoako ◽  
A. K. Gebeh ◽  
E. L. Marczylo ◽  
J. M. Willets ◽  
J. Elson ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Gene Selection ◽  
Cannabinoid Receptor ◽  
Receptor Gene ◽  
Expression Studies ◽  
Selection For ◽  
Gene Expression Studies ◽  
Receptor Gene Expression

scholarly journals Identification of stable reference genes for lipopolysaccharide-stimulated macrophage gene expression studies

2016 ◽  
Vol 1 (1) ◽  
Author(s):  
Roshini Kalagara ◽  
Weimin Gao ◽  
Honor L. Glenn ◽  
Colleen Ziegler ◽  
Laura Belmont ◽  
...  
Keyword(s):  
Gene Expression ◽  
Ribosomal Protein ◽  
Reference Gene ◽  
Reference Genes ◽  
Target Genes ◽  
Stable Gene ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Expression Studies ◽  
Gene Expression Studies

Gene expression studies which utilize lipopolysaccharide (LPS)-stimulated macrophages to model immune signaling are widely used for elucidating the mechanisms of inflammation-related disease. When expression levels of target genes are quantified using Real-Time quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR), they are analyzed in comparison to reference genes, which should have stable expression. Judicious selection of reference genes is, therefore, critical to interpretation of qRT-PCR results. Ideal reference genes must be identified for each experimental system and demonstrated to remain constant under the experimental conditions. In this study, we evaluated the stability of eight common reference genes: Beta-2-microglobulin (B2M), Cyclophilin A/Peptidylprolyl isomerase A, glyceraldehyde-3-phosphatedehydrogenase (GAPDH), Hypoxanthine Phosphoribosyltransferase 1, Large Ribosomal Protein P0, TATA box binding protein, Ubiquitin C (UBC), and Ribosomal protein L13A. Expression stability of each gene was tested under different conditions of LPS stimulation and compared to untreated controls. Reference gene stabilities were analyzed using Ct value comparison, NormFinder, and geNorm. We found that UBC, closely followed by B2M, is the most stable gene, while the commonly used reference gene GAPDH is the least stable. Thus, for improved accuracy in evaluating gene expression levels, we propose the use of UBC to normalize PCR data from LPS-stimulated macrophages.

scholarly journals Reference gene selection for head and neck squamous cell carcinoma gene expression studies

2009 ◽  
Vol 10 (1) ◽  
pp. 78 ◽  
Author(s):  
Benjamin Lallemant ◽  
Alexandre Evrard ◽  
Christophe Combescure ◽  
Heliette Chapuis ◽  
Guillaume Chambon ◽  
...  
Keyword(s):  
Gene Expression ◽  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Head And Neck ◽  
Reference Gene ◽  
Squamous Cell ◽  
Gene Selection ◽  
Expression Studies ◽  
Selection For ◽  
Gene Expression Studies

scholarly journals Current Practices for Reference Gene Selection in RT-qPCR of Aspergillus: Outlook and Recommendations for the Future

Genes ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 960
Author(s):  
Meagan Archer ◽  
Jianping Xu
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Reference Genes ◽  
Gene Selection ◽  
Quantitative Polymerase Chain Reaction ◽  
Specific Method ◽  
Experimental Conditions ◽  
Reference Gene Selection ◽  
Physiological Processes ◽  
The Stability

Aspergillus is a genus of filamentous fungi with vast geographic and ecological distributions. Species within this genus are clinically, agriculturally and biotechnologically relevant, leading to increasing interest in elucidating gene expression dynamics of key metabolic and physiological processes. Reverse-transcription quantitative Polymerase Chain Reaction (RT-qPCR) is a sensitive and specific method of quantifying gene expression. A crucial step for comparing RT-qPCR results between strains and experimental conditions is normalisation to experimentally validated reference gene(s). In this review, we provide a critical analysis of current reference gene selection and validation practices for RT-qPCR gene expression analyses of Aspergillus. Of 90 primary research articles obtained through our PubMed query, 17 experimentally validated the reference gene(s) used. Twenty reference genes were used across the 90 studies, with beta-tubulin being the most used reference gene, followed by actin, 18S rRNA and glyceraldehyde 3-phosphate dehydrogenase. Sixteen of the 90 studies used multiple reference genes for normalisation. Failing to experimentally validate the stability of reference genes can lead to conflicting results, as was the case for four studies. Overall, our review highlights the need to experimentally validate reference genes in RT-qPCR studies of Aspergillus.

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