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

Kód výsledku v 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>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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

Název v anglickém jazyce

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

Popis výsledku anglicky

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

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

GK - Lesnictví

OECD FORD obor

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Projekt

<a href="/cs/project/QH81039" target="_blank" >QH81039: Metody včasné identifikace, biologie, populační struktura, ochranná opatření a rozšíření karanténního škodlivého organismu Mycosphaerella pini (Dothistroma septospora) a dalších chorob asimilačního aparátu borovic v České republice</a><br>

Návaznosti

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

Rok uplatnění

2012

Kód důvěrnosti údajů

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

Název periodika

Journal of Agricultural Extension and Rural Development

ISSN

2141-2170

e-ISSN

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Svazek periodika

9

Číslo periodika v rámci svazku

4

Stát vydavatele periodika

NG - Nigérijská federativní republika

Počet stran výsledku

2

Strana od-do

221-222

Kód UT WoS článku

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EID výsledku v databázi Scopus

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