scholarly journals Identification of suitable reference genes for investigating gene expression in human gallbladder carcinoma using reverse transcription quantitative polymerase chain reaction

2014 ◽  
Vol 11 (4) ◽  
pp. 2967-2974 ◽  
Author(s):  
SHAN YU ◽  
QIWEI YANG ◽  
JING HUI YANG ◽  
ZHENWU DU ◽  
GUIZHEN ZHANG
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Gallbladder Carcinoma ◽  
Reverse Transcription ◽  
Reference Genes ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Human Gallbladder ◽  
Polymerase Chain ◽  
Suitable Reference

scholarly journals A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales (Delphinapterusleucas)

2017 ◽  
Author(s):  
Ming-An Tsai ◽  
I-Hua Chen ◽  
Jiann-Hsiung Wang ◽  
Shih-Jen Chou ◽  
Tsung-Hsien Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Immune System ◽  
Reverse Transcription ◽  
Reference Genes ◽  
Amplification Efficiency ◽  
Chain Reaction ◽  
Beluga Whales ◽  
Qrt Pcr ◽  
Polymerase Chain

Cytokines are fundamental for a functioning immune system, and thus, potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a novel probe-based quantitative gene expression assay for immunological assessment of captive beluga whales ( Delphinapterusleucas ) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNAlater solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset.

scholarly journals Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars

2018 ◽  
Vol 48 (11) ◽  
Author(s):  
Dennis Crystian ◽  
Jackeline Terto ◽  
José Vieira Silva ◽  
Cícero Almeida
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Drought Stress ◽  
Reference Genes ◽  
Quantitative Polymerase Chain Reaction ◽  
Elongation Factor ◽  
Chain Reaction ◽  
Stress Studies ◽  
Accurate Normalization ◽  
Polymerase Chain

ABSTRACT: The aim of this study was to analyze the expression of putative reference genes in sugarcane under drought stress. The varieties RB72454 and RB72910 were cultivated and the treatments control and drought stress applied to 135-day-old plants grown under field conditions. The stress level of the plants was measured by rate of photosynthesis, transpiration, and stomatal conductance. For each biological replicate, expression analyses were conducted using quantitative polymerase chain reaction for the genes α-tubulin, β-tubulin, β-actin, cyclophilin, eukaryotic elongation factor 1, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), histone H3 and ubiquitin. Stability was evaluated using the geNorm, NormFinder, and BestKeeper software packages. Among the candidate genes, GAPDH was identified as the most stable in all software, indicating its suitability for gene expression studies in sugarcane undergoing drought stress; the gene β-actin was the second most stable. These findings suggest using GAPDH and β-actin for normalization in relative gene expression in sugarcane.

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