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

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

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.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

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