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https://www.readbyqxmd.com/read/28598997/selection-of-reference-genes-for-quantitative-real-time-rt-pcr-assays-in-different-morphological-forms-of-dimorphic-zygomycetous-fungus-benjaminiella-poitrasii
#1
Ejaj K Pathan, Vandana Ghormade, Mukund V Deshpande
Benjaminiella poitrasii, a dimorphic non-pathogenic zygomycetous fungus, exhibits a morphological yeast (Y) to hypha (H) reversible transition in the vegetative phase, sporangiospores (S) in the asexual phase and zygospores (Z) in the sexual phase. To study the gene expression across these diverse morphological forms, suitable reference genes are required. In the present study, 13 genes viz. ACT, 18S rRNA, eEF1α, eEF-Tu,eIF-1A, Tub-α, Tub-b, Ubc, GAPDH, Try, WS-21, NADGDH and NADPGDH were evaluated for their potential as a reference, particularly for studying gene expression during the Y-H reversible transition and also for other asexual and sexual life stages of B...
2017: PloS One
https://www.readbyqxmd.com/read/28559905/validation-of-suitable-reference-genes-for-rt-qpcr-data-in-achyranthes-bidentata-blume-under-different-experimental-conditions
#2
Jinting Li, Xueping Han, Can Wang, Wanzhen Qi, Weiyu Zhang, Li Tang, Xiting Zhao
Real-time quantitative polymerase chain reaction (RT-qPCR) is a sensitive technique for gene expression studies. However, choosing the appropriate reference gene is essential to obtain reliable results for RT-qPCR assays. In the present work, the expression of eight candidate reference genes, EF1-α (elongation factor 1-α), GAPDH (glyceraldehyde 3-phosphate dehydrogenase), UBC (ubiquitin-conjugating enzyme), UBQ (polyubiquitin), ACT (actin), β-TUB (β-tubulin), APT1 (adenine phosphoribosyltransferase 1), and 18S rRNA (18S ribosomal RNA), was evaluated in Achyranthes bidentata samples using two algorithms, geNorm and NormFinder...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28523004/selection-of-suitable-reference-genes-for-quantitative-real-time-pcr-in-sapium-sebiferum
#3
Xue Chen, Yingji Mao, Shengwei Huang, Jun Ni, Weili Lu, Jinyan Hou, Yuting Wang, Weiwei Zhao, Minghao Li, Qiaojian Wang, Lifang Wu
Chinese tallow (Sapium sebiferum L.) is a promising landscape and bioenergy plant. Measuring gene expression by quantitative real-time polymerase chain reaction (qRT-PCR) can provide valuable information on gene function. Stably expressed reference genes for normalization are a prerequisite for ensuring the accuracy of the target gene expression level among different samples. However, the reference genes in Chinese tallow have not been systematically validated. In this study, 12 candidate reference genes (18S, GAPDH, UBQ, RPS15, SAND, TIP41, 60S, ACT7, PDF2, APT, TBP, and TUB) were investigated with qRT-PCR in 18 samples, including those from different tissues, from plants treated with sucrose and cold stresses...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28515733/stable-internal-reference-genes-for-normalizing-real-time-quantitative-pcr-in-baphicacanthus-cusia-under-hormonal-stimuli-and-uv-irradiation-and-in-different-plant-organs
#4
Yuxiang Huang, Hexin Tan, Jian Yu, Yue Chen, Zhiying Guo, Guoquan Wang, Qinglei Zhang, Junfeng Chen, Lei Zhang, Yong Diao
Baphicacanthus cusia (Nees) Bremek, the plant source for many kinds of drugs in traditional Chinese medicine, is widely distributed in South China, especially in Fujian. Recent studies about B. cusia mainly focus on its chemical composition and pharmacological effects, but further analysis of the plant's gene functions and expression is required to better understand the synthesis of its effective compounds. Real-time quantitative polymerase chain reaction (RT-qPCR) is a powerful method for gene expression analysis...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28453515/reliable-reference-genes-for-normalization-of-gene-expression-data-in-tea-plants-camellia-sinensis-exposed-to-metal-stresses
#5
Ming-Le Wang, Qing-Hui Li, Hua-Hong Xin, Xuan Chen, Xu-Jun Zhu, Xing-Hui Li
Tea plants [Camellia sinensis (L.) O. Kuntze] are an important leaf-type crop that are widely used for the production of non-alcoholic beverages in the world. Exposure to excessive amounts of heavy metals adversely affects the quality and yield of tea leaves. To analyze the molecular responses of tea plants to heavy metals, a reliable quantification of gene expression is important and of major importance herein is the normalization of the measured expression levels for the target genes. Ideally, stably expressed reference genes should be evaluated in all experimental systems...
2017: PloS One
https://www.readbyqxmd.com/read/28407308/methods-of-rna-preparation-affect-mrna-abundance-quantification-of-reference-genes-in-pig-maturing-oocytes
#6
Y-K Wang, X Li, Z-Q Song, C-X Yang
To ensure accurate normalization and quantification of target RNA transcripts using reverse transcription quantitative polymerase chain reaction (RT-qPCR), most studies focus on the identification of stably expressed gene(s) as internal reference. However, RNA preparation methods could also be an important factor, especially for test samples of limited quantity (e.g. oocytes). In this study, we aimed to select appropriate reference gene(s), and evaluate the effect of RNA preparation methods on gene expression quantification in porcine oocytes and cumulus cells during in vitro maturation...
April 13, 2017: Reproduction in Domestic Animals, Zuchthygiene
https://www.readbyqxmd.com/read/28362875/identification-and-validation-of-reference-genes-for-quantitative-real-time-pcr-studies-in-long-yellow-daylily-hemerocallis-citrina-borani
#7
Feifan Hou, Sen Li, Jinyao Wang, Xiuping Kang, Yiqun Weng, Guoming Xing
Gene expression analysis using reverse transcription quantitative real-time PCR (RT-qPCR) requires the use of reference gene(s) in the target species. The long yellow daylily, Hemerocallis citrina Baroni. is rich in beneficial secondary metabolites and is considered as a functional vegetable. It is widely cultivated and consumed in East Asian countries. However, reference genes for use in RT-qPCR in H. citrina are not available. In the present study, six potential reference genes, actin (ACT), AP-4 complex subunit (AP4), tubulin (TUB), ubiquitin (UBQ), 18S and 60S ribosomal RNA, were selected and their expression stability in different developmental stages, organs and accessions was evaluated using four statistical software packages (geNorm, NormFinder, BestKeeper, and RefFinder)...
2017: PloS One
https://www.readbyqxmd.com/read/28323834/selection-and-validation-of-reference-genes-for-qrt-pcr-analysis-during-biological-invasions-the-thermal-adaptability-of-bemisia-tabaci-med
#8
Tian-Mei Dai, Zhi-Chuang Lü, Wan-Xue Liu, Fang-Hao Wan
The Bemisia tabaci Mediterranean (MED) cryptic species has been rapidly invading to most parts of the world owing to its strong ecological adaptability, which is considered as a model insect for stress tolerance studies under rapidly changing environments. Selection of a suitable reference gene for quantitative stress-responsive gene expression analysis based on qRT-PCR is critical for elaborating the molecular mechanisms of thermotolerance. To obtain accurate and reliable normalization data in MED, eight candidate reference genes (β-act, GAPDH, β-tub, EF1-α, GST, 18S, RPL13A and α-tub) were examined under various thermal stresses for varied time periods by using geNorm, NormFinder and BestKeeper algorithms, respectively...
2017: PloS One
https://www.readbyqxmd.com/read/28067273/validation-of-reference-genes-for-cryopreservation-studies-with-the-gorgonian-coral-endosymbiont-symbiodinium
#9
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/27974876/evaluation-of-housekeeping-genes-for-quantitative-gene-expression-analysis-in-the-equine-kidney
#10
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
#11
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/27889881/identification-of-reference-genes-and-mirnas-for-qrt-pcr-in-human-esophageal-squamous-cell-carcinoma
#12
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/27835695/reference-genes-for-addressing-gene-expression-of-bladder-cancer-cell-models-under-hypoxia-a-step-towards-transcriptomic-studies
#13
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
https://www.readbyqxmd.com/read/27824922/identification-of-reference-genes-for-quantitative-real-time-pcr-in-date-palm-phoenix-dactylifera-l-subjected-to-drought-and-salinity
#14
Himanshu V Patankar, Dekoum V M Assaha, Rashid Al-Yahyai, Ramanjulu Sunkar, Mahmoud W Yaish
Date palm is an important crop plant in the arid and semi-arid regions supporting human population in the Middle East and North Africa. These areas have been largely affected by drought and salinity due to insufficient rainfall and improper irrigation practices. Date palm is a relatively salt- and drought-tolerant plant and more recently efforts have been directed to identifying genes and pathways that confer stress tolerance in this species. Quantitative real-time PCR (qPCR) is a promising technique for the analysis of stress-induced differential gene expression, which involves the use of stable reference genes for normalizing gene expression...
2016: PloS One
https://www.readbyqxmd.com/read/27803707/selection-of-reliable-reference-genes-for-gene-expression-studies-on-rhododendron-molle-g-don
#15
Zheng Xiao, Xiaobo Sun, Xiaoqing Liu, Chang Li, Lisi He, Shangping Chen, Jiale Su
The quantitative real-time polymerase chain reaction (qRT-PCR) approach has become a widely used method to analyze expression patterns of target genes. The selection of an optimal reference gene is a prerequisite for the accurate normalization of gene expression in qRT-PCR. The present study constitutes the first systematic evaluation of potential reference genes in Rhododendron molle G. Don. Eleven candidate reference genes in different tissues and flowers at different developmental stages of R. molle were assessed using the following three software packages: GeNorm, NormFinder, and BestKeeper...
2016: Frontiers in Plant Science
https://www.readbyqxmd.com/read/27761339/evaluation-of-reference-genes-for-gene-expression-in-red-tailed-phascogale-phascogale-calura-liver-lung-small-intestine-and-spleen
#16
Oselyne T W Ong, Lauren J Young, Julie M Old
BACKGROUND: Reference genes serve an important role as an endogenous control/standard for data normalisation in gene expression studies. Although reference genes have recently been suggested for marsupials, independent analysis of reference genes on different immune tissues is yet to be tested. Therefore, an assessment of reference genes is needed for the selection of stable, expressed genes across different marsupial tissues. METHODS: The study was conducted on red-tailed phascogales (Phascogale calura) using five juvenile and five adult males...
2016: PeerJ
https://www.readbyqxmd.com/read/27752147/identification-of-reliable-reference-genes-for-qrt-pcr-studies-of-the-developing-mouse-mammary-gland
#17
Anoeska Agatha Alida van de Moosdijk, Renée van Amerongen
Cell growth and differentiation are often driven by subtle changes in gene expression. Many challenges still exist in detecting these changes, particularly in the context of a complex, developing tissue. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) allows relatively high-throughput evaluation of multiple genes and developmental time points. Proper quantification of gene expression levels by qRT-PCR requires normalization to one or more reference genes. Traditionally, these genes have been selected based on their presumed "housekeeping" function, with the implicit assumption that they are stably expressed over the entire experimental set...
October 18, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27743995/withdrawn-selection-of-reference-genes-in-five-types-of-human-tissues-for-normalization-of-gene-expression-studies-in-infectious-diseases
#18
Cristina Silva Meira-Strejevitch, Vera Lucia Pereira-Chioccola, Marta Marques Maia, Daise Damaris Carnietto de Hipólito, Hui-Tzu Lin Wang, Gabriela Motoie, Aparecida Helena de Souza Gomes, Cristina Takami Kanamura, Roosecelis Brasil Martines, Cinara Cássia Brandão de Mattos, Fábio Batista Frederico, Luiz Carlos de Mattos, Cinara Cássia Brandão de Mattos, Fábio Batista Frederico, Rubens Camargo Siqueira, Mariana Previato, Amanda Pires Barbosa, Fernando Henrique Antunes Murata
This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.
October 12, 2016: Gene
https://www.readbyqxmd.com/read/27727313/endogenous-reference-genes-for-gene-expression-studies-on-bicuspid-aortic-valve-associated-aortopathy-in-humans
#19
Oliver J Harrison, Narain Moorjani, Christopher Torrens, Sunil K Ohri, Felino R Cagampang
Bicuspid aortic valve (BAV) disease is the most common congenital cardiac abnormality and predisposes patients to life-threatening aortic complications including aortic aneurysm. Quantitative real-time reverse transcription PCR (qRT-PCR) is one of the most commonly used methods to investigate underlying molecular mechanisms involved in aortopathy. The accuracy of the gene expression data is dependent on normalization by appropriate housekeeping (HK) genes, whose expression should remain constant regardless of aortic valve morphology, aortic diameter and other factors associated with aortopathy...
2016: PloS One
https://www.readbyqxmd.com/read/27521935/reference-gene-validation-for-quantification-of-gene-expression-during-final-oocyte-maturation-induced-by-diethylstilbestrol-and-di-2-ethylhexyl-phthalate-in-common-carp
#20
Yanyan Shi, Jie Lu, Yilei Wang, Shuhong Wang
Final oocyte maturation is the key step to successful spawning and fertilization. Quantitative real-time PCR (qPCR) is the technique of election to quantify the abundance of functional genes in such study. Reference gene is essential for correct interpretation of qPCR data. However, an ideal universal reference gene that is stable under all experimental circumstances has not been described. Researchers should validate their reference genes while performing qPCR analysis. The expression of 6 candidate reference genes: 18s rRNA, 28s rRNA, Cathepsin Z, Elongation factor 1-α, Glyceraldehyde-3-phosphate dehydrogenase and β-actin were investigated during final oocyte maturation induced by different compounds (DES and DEHP) in common carp (Cyprinus carpio)...
August 2016: Journal of Environmental Sciences (China)
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