Comparison of RNA isolation and associated methods for extracellular RNA detection by high-throughput quantitative polymerase chain reaction

2016 ◽  
Vol 501 ◽  
pp. 66-74 ◽  
Author(s):  
Kahraman Tanriverdi ◽  
Alper Kucukural ◽  
Ekaterina Mikhalev ◽  
Selim E. Tanriverdi ◽  
Rosalind Lee ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
High Throughput ◽  
Quantitative Polymerase Chain Reaction ◽  
Rna Isolation ◽  
Chain Reaction ◽  
Rna Detection ◽  
Extracellular Rna ◽  
Polymerase Chain

scholarly journals High-throughput applicable genomic sex typing of chicken by TaqMan real-time quantitative polymerase chain reaction

2010 ◽  
Vol 89 (7) ◽  
pp. 1451-1456 ◽  
Author(s):  
N.F. Rosenthal ◽  
H. Ellis ◽  
K. Shioda ◽  
C. Mahoney ◽  
K.R. Coser ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
High Throughput ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Sex Typing ◽  
Polymerase Chain

Gene Expression Profiling and Cancer-Related Pathways in Type I Endometrial Carcinoma

2010 ◽  
Vol 20 (5) ◽  
pp. 724-731 ◽  
Author(s):  
Fatma S.A. Saghir ◽  
Isa Mohamed Rose ◽  
Ahmad Zailani Hatta Mohd Dali ◽  
Zainab Shamsuddin ◽  
A Rahman A. Jamal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Normal Tissue ◽  
Transforming Growth Factor ◽  
Quantitative Polymerase Chain Reaction ◽  
Rna Isolation ◽  
Gene Set Enrichment Analysis ◽  
Type I ◽  
Chain Reaction ◽  
Polymerase Chain

Introduction:Malignant transformation of type I endometrium involves alteration in gene expression with subsequent uncontrolled proliferation of altered cells.Objective:The main objective of the present study was to identify the cancer-related genes and gene pathways in the endometrium of healthy and cancer patients.Materials and Methods:Thirty endometrial tissues from healthy and type I EC patients were subjected to total RNA isolation. The RNA samples with good integrity number were hybridized to a new version of Affymetrix Human Genome GeneChip 1.0 ST array. We analyzed the results using the GeneSpring 9.0 GX and the Pathway Studio 6.1 software. For validation assay, quantitative real-time polymerase chain reaction was used to analyze 4 selected genes in normal and EC tissue.Results:Of the 28,869 genes profiled, we identified 621 differentially expressed genes (2-fold) in the normal tissue and the tumor. Among these genes, 146 were up-regulated and 476 were down-regulated in the tumor as compared with the normal tissue (P < 0.001). Up-regulated genes included the v-erb-a erythroblastic leukemia viral oncogene homolog 3 (ErbB3), ErbB4, E74-like factor 3 (ELF3), and chemokine ligand 17 (CXCL17). The down-regulated genes included signal transducer and activator transcription 5B (STAT5b), transforming growth factor β receptor III (TGFβ3), caveolin 1 (CAV1), and protein kinase C alpha (PKCA). The gene set enrichment analysis showed 10 significant gene sets with related genes (P < 0.05). The quantitative polymerase chain reaction of 4 selected genes using similar RNA confirmed the microarray results (P < 0.05).Conclusions:Identification of molecular pathways with their genes related to type I EC contribute to the understanding of pathophysiology of this cancer, probably leading to identifying potential biomarkers of the cancer.

scholarly journals Development of a High-Throughput Method for Quantification of Plasmopara viticola DNA in Grapevine Leaves by Means of Quantitative Real-Time Polymerase Chain Reaction

2005 ◽  
Vol 95 (6) ◽  
pp. 672-678 ◽  
Author(s):  
G. Valsesia ◽  
D. Gobbin ◽  
A. Patocchi ◽  
A. Vecchione ◽  
I. Pertot ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
High Throughput ◽  
Fluorescent Dyes ◽  
Control Strategies ◽  
Quantitative Polymerase Chain Reaction ◽  
Plasmopara Viticola ◽  
Chain Reaction ◽  
High Throughput Method ◽  
Polymerase Chain

Plasmopara viticola is a strictly biotrophic oomycete that causes downy mildew, which is one of the most important grapevine diseases. Control of the disease is most often achieved by fungicide applications, which may have severe environmental consequences. Therefore, alternative control strategies based on biocontrol agents (BCAs) are currently in development. Thousands of potential BCAs have to be screened for their antagonist efficacy against Plasmopara viticola. Evaluation of their effect on the pathogen can be achieved by detecting the amount of P. viticola DNA in leaves treated with potential antagonists and infected with the pathogen. In this study, a rapid high-throughput method was developed for relative quantification of P. viticola DNA directly from Vitis vinifera leaves by means of multiplex real-time quantitative polymerase chain reaction (PCR) with TaqMan chemistry. This method allows simultaneous amplification, but independent detection, of pathogen and host DNA by using species-specific primers and TaqMan probes that are labeled with different fluorescent dyes. Including detection of V. vinifera DNA in the tests is fundamental because it provides an endogenous reference and allows normalization for variations caused by sample-to-sample differences in DNA extraction, PCR efficiencies, and pipetting volumes. The developed method allows highly sensitive and specific detection of P. viticola DNA (minimal detectable quantity of 0.1 pg). Moreover, high precision and reproducibility of TaqMan assays were observed over a linear range of four orders of magnitude, confirming the reliability of the developed PCR assay. Potential applications range from screening for BCA efficiency to evaluation of fungicide efficacy, or assessment of host resistance.

scholarly journals High-Throughput Quantitative Polymerase Chain Reaction in Picoliter Droplets

2008 ◽  
Vol 80 (23) ◽  
pp. 8975-8981 ◽  
Author(s):  
Margaret Macris Kiss ◽  
Lori Ortoleva-Donnelly ◽  
N. Reginald Beer ◽  
Jason Warner ◽  
Christopher G. Bailey ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
High Throughput ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Polymerase Chain
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