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https://www.readbyqxmd.com/read/29166265/selection-of-reference-genes-for-quantitative-real-time-rt-pcr-on-gene-expression-in-golden-pompano-trachinotus-ovatus
#1
X J Chen, X Q Zhang, S Huang, Z J Cao, Q W Qin, W T Hu, Y Sun, Y C Zhou
Golden pompano (Trachinotus ovatus) is an important economically fish species. In this study, with an aim to identify reliable reference genes for quantitative real-time PCR (qRT-PCR) in golden pompano, we evaluated the expression stability of eight housekeeping genes in the presence and absence of poly I:C stimulation in eight tissues. The PCR data was analyzed by geNorm and NormFinder algorithms. The results showed that the expression of all the examined genes exhibited tissue-dependent variations. When under normal physiological condition, geNorm and NormFinder identified B2M and 18S as suitable genes...
September 26, 2017: Polish Journal of Veterinary Sciences
https://www.readbyqxmd.com/read/29134774/identifying-suitable-reference-genes-for-developing-and-injured-mouse-cns-tissues
#2
Dongchao Xu, Ajuan Liu, Xuan Wang, Ming Zhang, Zunyi Zhang, Tan Zhou, Mengsheng Qiu
Accurate quantification of gene expression is fundamental for understanding the molecular, genetic and functional bases of tissue development and diseases. Quantitative real-time PCR (qPCR) is now the most widely used method of quantifying gene expression due to its simplicity, specificity, sensitivity and wide quantification range. The use of appropriate reference genes to ensure accurate normalization is crucial for the correct quantification of gene expression from the early development, maturation, aging to injury processes in the central nervous system (CNS)...
November 14, 2017: Developmental Neurobiology
https://www.readbyqxmd.com/read/29091948/selection-and-validation-of-appropriate-reference-genes-for-quantitative-real-time-pcr-analysis-in-salvia-hispanica
#3
Rahul Gopalam, Sunny D Rupwate, Ajay W Tumaney
Quantitative real-time polymerase chain reaction (qRT-PCR) has become the most popular choice for gene expression studies. For accurate expression analysis, it is pertinent to select a stable reference gene to normalize the data. It is now known that the expression of internal reference genes varies considerably during developmental stages and under different experimental conditions. For Salvia hispanica, an economically important oilseed crop, there are no reports of stable reference genes till date. In this study, we chose 13 candidate reference genes viz...
2017: PloS One
https://www.readbyqxmd.com/read/29036603/identification-of-optimal-reference-genes-for-transcriptomic-analyses-in-normal-and-diseased-human-heart
#4
Cristina E Molina, Eric Jacquet, Prishila Ponien, Christian Muñoz-Guijosa, Istvan Baczkó, Lars S Maier, Patrick Donzeau-Gouge, Dobromir Dobrev, Rodolphe Fischmeister, Anne Garnier
Aims: Quantitative real-time RT-PCR (RT-qPCR) has become the method of choice for mRNA quantification, but requires an accurate normalization based on the use of reference genes showing invariant expression across various pathological conditions. Only few data exist on appropriate reference genes for the human heart. The objective of this study was to determine a set of suitable reference genes in human atrial and ventricular tissues, from right and left cavities in control and in cardiac diseases...
September 22, 2017: Cardiovascular Research
https://www.readbyqxmd.com/read/28970964/identification-of-reference-genes-in-blood-before-and-after-entering-the-plateau-for-sybr-green-rt-qpcr-studies
#5
Jun Xiao, Xiaowei Li, Juan Liu, Xiu Fan, Huifen Lei, Cuiying Li
BACKGROUND: Tibetans have lived at high altitudes for thousands of years, and they have unique physiological traits that enable them to tolerate this hypoxic environment. However, the genetic basis of these traits is still unknown. As a sensitive and highly efficient technique, RT-qPCR is widely used in gene expression analyses to provide insight into the molecular mechanisms underlying environmental changes. However, the quantitative analysis of gene expression in blood is limited by a shortage of stable reference genes for the normalization of mRNA levels...
2017: PeerJ
https://www.readbyqxmd.com/read/28964897/comprehensive-evaluation-of-candidate-reference-genes-for-gene-expression-studies-in-lysiphlebia-japonica-hymenoptera-aphidiidae-using-rt-qpcr
#6
Xue-Ke Gao, Shuai Zhang, Jun-Yu Luo, Chun-Yi Wang, Li-Min Lü, Li-Juan Zhang, Xiang-Zhen Zhu, Li Wang, Hui Lu, Jin-Jie Cui
Lysiphlebia japonica (Ashmead) is a predominant parasitoid of cotton-melon aphids in the fields of northern China with a proven ability to effectively control cotton aphid populations in early summer. For accurate normalization of gene expression in L. japonica using quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR), reference genes with stable gene expression patterns are essential. However, no appropriate reference genes is L. japonica have been investigated to date. In the present study, 12 selected housekeeping genes from L...
September 27, 2017: Gene
https://www.readbyqxmd.com/read/28946009/selection-of-reference-genes-for-quantitative-real-time-pcr-analysis-in-chicken-ovary-following-silver-nanoparticle-treatment
#7
Dorota Katarzyńska-Banasik, Małgorzata Grzesiak, Andrzej Sechman
Reliable results of quantitative real time PCR (qPCR) analysis require normalization of target gene expression level using reference genes (RGs). However housekeeping genes expression may vary under experimental conditions, so selection of the proper RGs is a crucial step in a qPCR analysis. Several algorithms have been developed to address this problem: geNorm, NormFinder and BestKeeper. In this study, we have used these three tools to evaluate the stability of RGs in the ovarian tissues of hens treated with silver nanoparticles...
September 20, 2017: Environmental Toxicology and Pharmacology
https://www.readbyqxmd.com/read/28919905/candidate-reference-genes-selection-and-application-for-rt-qpcr-analysis-in-kenaf-with-cytoplasmic-male-sterility-background
#8
Bujin Zhou, Peng Chen, Aziz Khan, Yanhong Zhao, Lihong Chen, Dongmei Liu, Xiaofang Liao, Xiangjun Kong, Ruiyang Zhou
Cytoplasmic male sterility (CMS) is a maternally inherited trait that results in the production of dysfunctional pollen. Based on reliable reference gene-normalized real-time quantitative PCR (RT-qPCR) data, examining gene expression profile can provide valuable information on the molecular mechanism of kenaf CMS. However, studies have not been conducted regarding selection of reference genes for normalizing RT-qPCR data in the CMS and maintainer lines of kenaf crop. Therefore, we studied 10 candidate reference genes (ACT3, ELF1A, G6PD, PEPKR1, TUB, TUA, CYP, GAPDH, H3, and 18S) to assess their expression stability at three stages of pollen development in CMS line 722A and maintainer line 722B of kenaf...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28810828/suitable-reference-gene-for-quantitative-real-time-pcr-analysis-of-gene-expression-in-gonadal-tissues-of-minnow-puntius-sophore-under-high-temperature-stress
#9
Arabinda Mahanty, Gopal Krishna Purohit, Sasmita Mohanty, Nihar Ranjan Nayak, Bimal Prasanna Mohanty
BACKGROUND: High ambient temperature is known to affect fish gonadal development and physiology in a variety of ways depending on the severity and duration of exposure; however, the underlying molecular mechanisms are poorly understood. Gonadal gene expression influence the gonadal development, physiology and the quality of egg/sperm produced in teleosts and the mechanistic understanding of spatio-temporal changes in the gonadal gene expression could be instrumental in controlling the fate of egg/sperm and the quality of seed produced...
August 15, 2017: BMC Genomics
https://www.readbyqxmd.com/read/28763741/expression-stability-and-selection-of-optimal-reference-genes-for-gene-expression-normalization-in-early-life-stage-rainbow-trout-exposed-to-cadmium-and-copper
#10
Kamran Shekh, Song Tang, Som Niyogi, Markus Hecker
Gene expression analysis represents a powerful approach to characterize the specific mechanisms by which contaminants interact with organisms. One of the key considerations when conducting gene expression analyses using quantitative real-time reverse transcription-polymerase chain reaction (qPCR) is the selection of appropriate reference genes, which is often overlooked. Specifically, to reach meaningful conclusions when using relative quantification approaches, expression levels of reference genes must be highly stable and cannot vary as a function of experimental conditions...
September 2017: Aquatic Toxicology
https://www.readbyqxmd.com/read/28746411/selection-and-validation-of-reference-genes-for-quantitative-real-time-pcr-analysis-under-different-experimental-conditions-in-the-leafminer-liriomyza-trifolii-diptera-agromyzidae
#11
Ya-Wen Chang, Jing-Yun Chen, Ming-Xing Lu, Yuan Gao, Zi-Hua Tian, Wei-Rong Gong, Wei Zhu, Yu-Zhou Du
Liriomyza trifolii is a highly-invasive leafmining insect that causes significant damage to vegetables and horticultural crops worldwide. Relatively few studies have quantified gene expression in L. trifolii using real-time quantitative PCR (RT-qPCR), which is a reliable and sensitive technique for measuring gene expression. RT-qPCR requires the selection of reference genes to normalize gene expression data and control for internal differences between samples. In this study, nine housekeeping genes from L. trifolii were selected for their suitability in normalizing gene expression using geNorm, Normfinder, BestKeeper, the ΔCt method and RefFinder...
2017: PloS One
https://www.readbyqxmd.com/read/28699725/evaluation-of-candidate-reference-genes-for-quantitative-expression-studies-in-asian-seabass-lates-calcarifer-during-ontogenesis-and-in-tissues-of-healthy-and-infected-fishes
#12
Anutosh Paria, Jie Dong, P P Suresh Babu, M Makesh, Aparna Chaudhari, A R Thirunavukkarasu, C S Purushothaman, K V Rajendran
Quantitative real-time PCR (qRT-PCR), used to determine the gene expression profile, is an important tool in functional genomic research, including fishes. To obtain more robust and meaningful result, the best possible normalization of the data is of utmost significance. In the present study, we have evaluated the potential of five commonly used housekeeping genes i.e., elongation factor 1-α (EF1A), β-Actin (ACTB), 18S ribosomal RNA (18S), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-2-Microglobulin (B2M) in normal physiological conditions, developmental stages and in response to bacterial infection in Asian seabass, Lates calcarifer (Bloch), an important food fish cultured in the Asia-Pacific region...
September 2016: Indian Journal of Experimental Biology
https://www.readbyqxmd.com/read/28671969/reference-gene-selection-for-the-shell-gland-of-laying-hens-in-response-to-time-points-of-eggshell-formation-and-nicarbazin
#13
Sami Samiullah, Juliet Roberts, Shu-Biao Wu
Ten reference genes were investigated for normalization of gene expression data in the shell gland of laying hens. Analyses performed with geNorm revealed that hypoxanthine phosphoribosyltransferase 1 (HPRT1) and hydroxymethylbilane synthase (HMBS) were the two most stable reference genes in response to post-oviposition time alone (POT) or with nicarbazin treatment (POT+N) of laying hens. NormFinder analyses showed that the two most stable reference genes in response to POT and POT+N were 18S ribosomal RNA (18S rRNA), ribosomal protein L4 (RPL4) and HMBS, RPL4, respectively...
2017: PloS One
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
#14
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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