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https://www.readbyqxmd.com/read/28090788/selection-of-suitable-reference-genes-for-quantitative-real-time-pcr-in-trabecular-meshwork-cells-under-oxidative-stress
#1
Jing Zhao, HongYan Zhou, Lixia Sun, Ben Yang, Lin Zhang, Hongfeng Shi, Yajuan Zheng
Oxidative stress-induced dysfunction in trabecular meshwork (TM) cells is considered a major alteration that can lead to glaucoma. H2O2 is the most widely used agent for inducing oxidation in TM cells in vitro. Quantitative real-time PCR (qPCR) is an important method for studying alterations in gene expression, and suitable (i.e., invariant) reference genes must be defined to normalize expression levels. In this study, eight common reference genes, i.e., PRS18, ACTB, B2M, GAPDH, PPIA, HPRT1, YWHAZ, and TBP, were evaluated for use in studies of H2O2-induced dysfunction in TM cells...
January 15, 2017: Free Radical Research
https://www.readbyqxmd.com/read/28067273/validation-of-reference-genes-for-cryopreservation-studies-with-the-gorgonian-coral-endosymbiont-symbiodinium
#2
Gabriella Chong, Fu-Wen Kuo, Sujune Tsai, Chiahsin Lin
Quantification by real-time RT-PCR requires a stable internal reference known as a housekeeping gene (HKG) for normalising the mRNA levels of target genes. The present study identified and validated stably expressed HKGs in post-thaw Symbiodinium clade G. Six potential HKGs, namely, pcna, gapdh, 18S rRNA, hsp90, rbcl, and ps1, were analysed using three different algorithms, namely, GeNorm, NormFinder, and BestKeeper. The GeNorm algorithm ranked the candidate genes as follows in the order of decreasing stability: pcna and gapdh > ps1 > 18S rRNA > hsp90 > rbcl...
January 9, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28056110/selection-of-reference-genes-for-qrt-pcr-analysis-of-gene-expression-in-stipa-grandis-during-environmental-stresses
#3
Dongli Wan, Yongqing Wan, Qi Yang, Bo Zou, Weibo Ren, Yong Ding, Zhen Wang, Ruigang Wang, Kai Wang, Xiangyang Hou
Stipa grandis P. Smirn. is a dominant plant species in the typical steppe of the Xilingole Plateau of Inner Mongolia. Selection of suitable reference genes for the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) is important for gene expression analysis and research into the molecular mechanisms underlying the stress responses of S. grandis. In the present study, 15 candidate reference genes (EF1 beta, ACT, GAPDH, SamDC, CUL4, CAP, SNF2, SKIP1, SKIP5, SKIP11, UBC2, UBC15, UBC17, UCH, and HERC2) were evaluated for their stability as potential reference genes for qRT-PCR under different stresses...
2017: PloS One
https://www.readbyqxmd.com/read/28050855/selection-of-reference-genes-from-shiraia-bambusicola-for-rt-qpcr-analysis-under-different-culturing-conditions
#4
Chen Zhang, Tong Li, Cheng-Lin Hou, Xiao-Ye Shen
Stable reference genes are necessary to analyse quantitative real-time reverse transcription PCR (qRT-PCR) data and determine the reliability of the final results. For further studies of the valuable fungus Shiraia bambusicola, the identification of suitable reference genes has become increasingly urgent. In this study, three conventional reference genes and nine novel candidates were evaluated under different light conditions (all-dark, all-light and 12-h light/dark) and in different media (rice medium, PD medium, and Czapek-Dox medium)...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28046130/evaluation-of-reference-genes-for-normalization-of-gene-expression-using-quantitative-rt-pcr-under-aluminum-cadmium-and-heat-stresses-in-soybean
#5
Mengmeng Gao, Yaping Liu, Xiao Ma, Qin Shuai, Junyi Gai, Yan Li
Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is widely used to analyze the relative gene expression level, however, the accuracy of qRT-PCR is greatly affected by the stability of reference genes, which is tissue- and environment- dependent. Therefore, choosing the most stable reference gene in a specific tissue and environment is critical to interpret gene expression patterns. Aluminum (Al), cadmium (Cd), and heat stresses are three important abiotic factors limiting soybean (Glycine max) production in southern China...
2017: PloS One
https://www.readbyqxmd.com/read/28040059/validation-of-housekeeping-genes-for-the-normalization-of-rt-qpcr-expression-studies-in-oral-squamous-cell-carcinoma-cell-line-treated-by-5-kinds-of-chemotherapy-drugs
#6
W Song, W H Zhang, H Zhang, Y Li, Y Zhang, W Yin, Q Yang
Reverse transcription quantitative polymerase chain reaction (RT-qPCR) has become a frequently used strategy in gene expression studies. The relative quantification method is an important and commonly used method for the evaluation of RT-qPCR data. The key of this method is to identify an applicable internal control gene because the usage of different internal control genes may lead to distinct conclusions. Herein, we report the validation of 12 common housekeeping genes for RT-qPCR for gene expression analysis in the Oral squamous cell carcinoma (OSCC) cell line (KB and Tca-8113) treated by 5 kinds of Chemotherapy Drugs...
November 30, 2016: Cellular and Molecular Biology
https://www.readbyqxmd.com/read/28039034/development-and-evaluation-of-a-novel-rt-qpcr-based-test-for-the-quantification-of-her2-gene-expression-in-breast-cancer
#7
Hicham El Hadi, Imane Abdellaoui-Maane, Denise Kottwitz, Manal El Amrani, Nadia Bouchoutrouch, Zineb Qmichou, Mehdi Karkouri, Hicham ElAttar, Hassan Errihani, Pedro L Fernandez, Youssef Bakri, Hassan Sefrioui, Abdeladim Moumen
Accurate measurement of Human epidermal growth factor receptor (HER2) gene expression is central for breast or stomach cancer therapy orientation and prognosis. The current standards testing methods for HER2 expression are immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). In the current study, we explored the use of quantitative real time reverse transcription-PCR (RT-qPCR) as a potential method for the accurate relative quantification of the HER2 gene using formalin fixed paraffin embedded (FFPE) breast cancer biopsy samples...
December 28, 2016: Gene
https://www.readbyqxmd.com/read/28036291/assessing-the-clinical-value-of-micrornas-in-formalin-fixed-paraffin-embedded-liposarcoma-tissues-overexpressed-mir-155-is-an-indicator-of-poor-prognosis
#8
Nikolaos Kapodistrias, Konstantinos Mavridis, Anna Batistatou, Penelope Gogou, Vasilios Karavasilis, Ioannis Sainis, Evangelos Briasoulis, Andreas Scorilas
Liposarcoma (LPS) is a malignancy with extreme heterogeneity and thus optimization towards personalizing patient prognosis and treatment is essential. Here, we evaluated miR-155, miR-21, miR-143, miR-145 and miR-451 that are implicated in LPS, as novel FFPE tissue biomarkers.A total of 83 FFPE tissue specimens from primary LPS and lipomas (LPM) were analyzed. A proteinase K incubation-Trizol treatment coupled protocol was used for RNA isolation. After polyadenylation of total RNA and reverse transcription, expression analysis of 9 candidate reference and 5 target miRNAs was performed by qPCR...
December 28, 2016: Oncotarget
https://www.readbyqxmd.com/read/27994956/identifying-suitable-reference-genes-for-gene-expression-analysis-in-developing-skeletal-muscle-in-pigs
#9
Guanglin Niu, Yalan Yang, YuanYuan Zhang, Chaoju Hua, Zishuai Wang, Zhonglin Tang, Kui Li
The selection of suitable reference genes is crucial to accurately evaluate and normalize the relative expression level of target genes for gene function analysis. However, commonly used reference genes have variable expression levels in developing skeletal muscle. There are few reports that systematically evaluate the expression stability of reference genes across prenatal and postnatal developing skeletal muscle in mammals. Here, we used quantitative PCR to examine the expression levels of 15 candidate reference genes (ACTB, GAPDH, RNF7, RHOA, RPS18, RPL32, PPIA, H3F3, API5, B2M, AP1S1, DRAP1, TBP, WSB, and VAPB) in porcine skeletal muscle at 26 different developmental stages (15 prenatal and 11 postnatal periods)...
2016: PeerJ
https://www.readbyqxmd.com/read/27976798/identification-and-validation-of-micrornas-as-endogenous-controls-for-quantitative-polymerase-chain-reaction-in-plasma-for-stable-coronary-artery-disease
#10
Yuejuan Zhang, Wenxian Tang, Ling Peng, Jinqiang Tang, Zhaokai Yuan
BACKGROUND: Circulating microRNAs (miRNAs) have been proved to serve as biomarkers for diagnosis and assessment of prognosis of coronary artery disease (CAD). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a widely-used technique to estimate expression levels of circulating miRNAs. Selection of optimal endogenous control (EC) remains critical to obtain reliable qPCR data of miRNAs expression. However, reference controls for normalization of circulating miRNA in CAD are still lacking...
2016: Cardiology Journal
https://www.readbyqxmd.com/read/27974876/evaluation-of-housekeeping-genes-for-quantitative-gene-expression-analysis-in-the-equine-kidney
#11
Sara Azarpeykan, Keren E Dittmer
Housekeeping genes (HKGs) are used as internal controls for normalising and calculating the relative expression of target genes in RT-qPCR experiments. There is no unique universal HKG and HKGs vary among organisms and tissues, so this study aimed to determine the most stably expressed HKGs in the equine kidney. The evaluated HKGs included 18S ribosomal RNA (18S), 28S ribosomal RNA (28S), ribosomal protein L32 (RPL32), β-2-microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), succinate dehydrogenase complex (SDHA), zeta polypeptide (YWHAZ), and hypoxanthine phosphoribosyltransferase 1 (HPRT1)...
2016: Journal of Equine Science
https://www.readbyqxmd.com/read/27957397/validation-of-internal-reference-genes-for-relative-quantitation-studies-of-gene-expression-in-human-laryngeal-cancer
#12
Xiaofeng Wang, Jinting He, Wei Wang, Ming Ren, Sujie Gao, Guanjie Zhao, Jincheng Wang, Qiwei Yang
BACKGROUND: The aim of this study was to determine the expression stabilities of 12 common internal reference genes for the relative quantitation analysis of target gene expression performed by reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) in human laryngeal cancer. METHODS: Hep-2 cells and 14 laryngeal cancer tissue samples were investigated. The expression characteristics of 12 internal reference gene candidates (18S rRNA, GAPDH, ACTB, HPRT1, RPL29, HMBS, PPIA, ALAS1, TBP, PUM1, GUSB, and B2M) were assessed by RT-qPCR...
2016: PeerJ
https://www.readbyqxmd.com/read/27942007/validation-of-reference-genes-for-real-time-quantitative-pcr-qpcr-analysis-of-avibacterium-paragallinarum
#13
Shuxiang Wen, Xiaoling Chen, Fuzhou Xu, Huiling Sun
Real-time quantitative reverse transcription PCR (qRT-PCR) offers a robust method for measurement of gene expression levels. Selection of reliable reference gene(s) for gene expression study is conducive to reduce variations derived from different amounts of RNA and cDNA, the efficiency of the reverse transcriptase or polymerase enzymes. Until now reference genes identified for other members of the family Pasteurellaceae have not been validated for Avibacterium paragallinarum. The aim of this study was to validate nine reference genes of serovars A, B, and C strains of A...
2016: PloS One
https://www.readbyqxmd.com/read/27922100/evaluation-of-candidate-reference-genes-for-rt-qpcr-studies-in-three-metabolism-related-tissues-of-mice-after-caloric-restriction
#14
Huan Gong, Liang Sun, Beidong Chen, Yiwen Han, Jing Pang, Wei Wu, Ruomei Qi, Tie-Mei Zhang
Reverse transcription quantitative-polymerase chain reaction (RT-qPCR) is a routine method for gene expression analysis, and reliable results depend on proper normalization by stable reference genes. Caloric restriction (CR) is a robust lifestyle intervention to slow aging and delay onset of age-associated diseases via inducing global changes in gene expression. Reliable normalization of RT-qPCR data becomes crucial in CR studies. In this study, the expression stability of 12 candidate reference genes were evaluated in inguinal white adipose tissue (iWAT), skeletal muscle (Sk...
December 6, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27889881/identification-of-reference-genes-and-mirnas-for-qrt-pcr-in-human-esophageal-squamous-cell-carcinoma
#15
Li Chen, Yulin Jin, Lin Wang, Fenghao Sun, Xiaodong Yang, Mengkun Shi, Cheng Zhan, Yu Shi, Qun Wang
It is important to select an appropriate reference gene and miRNA when using quantitative real-time polymerase chain reaction (qRT-PCR) to analyze gene and miRNA expression. However, many commonly used reference genes and miRNAs are not stably expressed and therefore not suitable for normalization or quantification of qRT-PCR data. This study aims to identify appropriate reference genes and miRNAs for use in human esophageal squamous carcinoma qRT-PCR analysis. Using data provided by The Cancer Genome Atlas, we identified DDX5, LAPTM4A, P4HB, RHOA, miR-28-5p, miR-34a-5p, and miR-186-5p as candidate reference genes and miRNAs...
January 2017: Medical Oncology
https://www.readbyqxmd.com/read/27878453/evaluation-of-stability-and-validation-of-reference-genes-for-rt-qpcr-expression-studies-in-rice-plants-under-water-deficit
#16
Priscila Ariane Auler, Letícia Carvalho Benitez, Marcelo Nogueira do Amaral, Isabel Lopes Vighi, Gabriela Dos Santos Rodrigues, Luciano Carlos da Maia, Eugenia Jacira Bolacel Braga
Many studies use strategies that allow for the identification of a large number of genes expressed in response to different stress conditions to which the plant is subjected throughout its cycle. In order to obtain accurate and reliable results in gene expression studies, it is necessary to use reference genes, which must have uniform expression in the majority of cells in the organism studied. RNA isolation of leaves and expression analysis in real-time quantitative polymerase chain reaction (RT-qPCR) were carried out...
November 23, 2016: Journal of Applied Genetics
https://www.readbyqxmd.com/read/27878262/identification-of-suitable-reference-genes-for-bdv-infected-primary-rat-hippocampal-neurons
#17
Qiang Mao, Lujun Zhang, Yujie Guo, Lu Sun, Siwen Liu, Peng He, Rongzhong Huang, Lin Sun, Shigang Chen, Hong Zhang, Peng Xie
Borna disease virus (BDV) is a neurotropic RNA virus that infects the limbic system of mammals and results in behavioral disorders. The hippocampus is a core region in the limbic system, which contributes to memory and learning and is important in the regulation of emotion. However, no validated microRNA housekeeping genes have yet been identified in BDV‑infected rat primary hippocampal neurons. Proper normalization is key in accurate miRNA expression analysis. The present study used reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) to evaluate the expression stability of 10 commonly used reference genes [miR‑92a, 5S, U6, miR‑103, miR‑101a, miR-let-7a, miR‑16, E2 small nucleolar RNA (snoRNA), U87 and miR‑191] in BDV‑infected rat hippocampal neurons and non‑infected controls across 12 days post‑infection...
December 2016: Molecular Medicine Reports
https://www.readbyqxmd.com/read/27877186/selection-of-reference-genes-for-rt-qpcr-analysis-in-coccinella-septempunctata-to-assess-un-intended-effects-of-rnai-transgenic-plants
#18
Chunxiao Yang, Evan L Preisser, Hongjun Zhang, Yong Liu, Liangying Dai, Huipeng Pan, Xuguo Zhou
The development of genetically engineered plants that employ RNA interference (RNAi) to suppress invertebrate pests opens up new avenues for insect control. While this biotechnology shows tremendous promise, the potential for both non-target and off-target impacts, which likely manifest via altered mRNA expression in the exposed organisms, remains a major concern. One powerful tool for the analysis of these un-intended effects is reverse transcriptase-quantitative polymerase chain reaction, a technique for quantifying gene expression using a suite of reference genes for normalization...
2016: Frontiers in Plant Science
https://www.readbyqxmd.com/read/27869224/selection-of-reliable-reference-genes-for-normalization-of-quantitative-rt-pcr-from-different-developmental-stages-and-tissues-in-amphioxus
#19
Qi-Lin Zhang, Qian-Hua Zhu, Xin Liao, Xiu-Qiang Wang, Tao Chen, Han-Ting Xu, Juan Wang, Ming-Long Yuan, Jun-Yuan Chen
Amphioxus is a closest living proxy to the ancestor of cephalochordates with vertebrates, and key animal for novel understanding in the evolutionary origin of vertebrate body plan, genome, tissues and immune system. Reliable analyses using quantitative real-time PCR (qRT-PCR) for answering these scientific questions is heavily dependent on reliable reference genes (RGs). In this study, we evaluated stability of thirteen candidate RGs in qRT-PCR for different developmental stages and tissues of amphioxus by four independent (geNorm, NormFinder, BestKeeper and deltaCt) and one comparative algorithms (RefFinder)...
November 21, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27835695/reference-genes-for-addressing-gene-expression-of-bladder-cancer-cell-models-under-hypoxia-a-step-towards-transcriptomic-studies
#20
Luís Lima, Cristiana Gaiteiro, Andreia Peixoto, Janine Soares, Manuel Neves, Lúcio Lara Santos, José Alexandre Ferreira
Highly aggressive, rapidly growing tumors contain significant areas of hypoxia or anoxia as a consequence of inadequate and/or irregular blood supply. During oxygen deprivation, tumor cells withstand a panoply of adaptive responses, including a shift towards anaerobic metabolism and the reprogramming of the transcriptome. One of the major mediators of the transcriptional hypoxic response is the hypoxia-inducible factor 1 (HIF-1), whose stabilization under hypoxia acts as an oncogenic stimulus contributing to chemotherapy resistance, invasion and metastasis...
2016: PloS One
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