Quantification of Dothistroma septosporum spores by real-time polymerase chain reaction (PCR)

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62156489%3A43410%2F12%3A00194173" target="_blank" >RIV/62156489:43410/12:00194173 - isvavai.cz</a>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Quantification of Dothistroma septosporum spores by real-time polymerase chain reaction (PCR)

Original language description

Air sampling can be combined with modern sensitive molecular methods such as quantitative polymerase chain reaction (qPCR), real-time PCR. A method for absolute quantification of Dothistroma septosporum (teleomorph Mycosphaerella pini) spores by real-time PCR is being developed. We report here the optimisation of plasmid standards and standard curves that are required for quantification using qPCR. Preparation of a DNA standard is necessary to make a robust and reliable estimation of the number of spores. The most consistent results have been obtained by using plasmid DNA with insert containing target sequence (Dhanasekaran et al., 2010). The standard can be used to determine the number of DNA copies in a spore trap sample, and thus the number of spores. Two plasmids, with different lengths of the target gene inserted (single copy gene), were prepared and both linear and circular forms of these were used for standard curve generation. Linear plasmid amplified in average of 2 to 3.5 cyc

Czech name

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Czech description

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Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

GK - Forestry

OECD FORD branch

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Project

<a href="/en/project/QH81039" target="_blank" >QH81039: Methods of early identification, biology, genetic structure, control and distribution of Dothistroma needle blight and other diseases of pines in the Czech Republic</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2012

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Agricultural Extension and Rural Development

ISSN

2141-2170

e-ISSN

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Volume of the periodical

9

Issue of the periodical within the volume

4

Country of publishing house

NG - NIGERIA

Number of pages

2

Pages from-to

221-222

UT code for WoS article

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EID of the result in the Scopus database

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