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The use of microsatellite analysis for the characterisation of onion
Mitrová, Katarína ; Ovesná, Jaroslava
The object of the methodology is the process characterizing the autenticity of varieties of onion put through microsatellite analysis (SSR- Single Sequence Repeats) for the characterization of varieties of garlic into the practice. This method allows to identify varieties of onion and characterize genetic resources using DNA markers and determine their genetic similarity based on the length variability of short repeating sequences (microsatellites) in the characterization of varieties of onion, Allium cepa L. The principle of the method is amplification of the genome containing the microsatelite locus by polymerase chain reaction (PCR) using specific primers and subsequent analysis of the lengtht of the products and the identification of the DNA profile to the declared variety.
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The use of microsatellite analysis for the characterisation of garlic
Mitrová, Katarína ; Ovesná, Jaroslava ; Svobodová, Leona
The object of the methodology is the process characterizing the autenticity of varieties of garlic put through microsatellite analysis (SSR- Single Sequence Repeats) for the characterization of varieties of garlic into the practice. This method allows to identify varieties of garlic and characterize genetic resources using DNA markers and determine their genetic similarity based on the length variability of short repeating sequences (microsatellites) in the characterization of varieties of garlic, Allium sativum L. The principle of the method is amplification of the genome containing the microsatelite locus by polymerase chain reaction (PCR) using specific primers and subsequent analysis of the lengtht of the products and the identification of the DNA profile to the declared variety.
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Methodology for analysis of dairy products and their soy analogues for the presence of RoundUp Ready soybean
Tesařová, Zuzana ; Šídová, Dana ; Vráblík, Aleš ; Hodek, Jan ; Ovesná, Jaroslava
Dairy products may be subject to adulteration, where there is a relatively expensive raw material (e.g. cream) replaced by the addition of vegetable fats. Milk can be replaced in dairy products for example by soybean extract. Because soy belongs among strong allergens and the incidence of allergy is rather high. Therefore it is necessary to have a method for its detection. This work introduces the method for detection of soy, including analysis of genetic modified RoundUp Ready soybean. Detection of soybean in dairy products is based on analysis of amplification products melting profiles by High Resolution Melting (HRM) method. Analysis of geneticaly modified RoundUp Ready soybean is based on DNA detection via Polymerase Chain Reaction(PCR).
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Zabezpečení laboratorní detekce geneticky modifikovaných organismů
Výzkumný ústav rostlinné výroby‚ Praha-Ruzyně ; Koucká, Petra ; Hodek, Jan ; Pouchová, Vladimíra ; Kučera, Ladislav ; Ovesná, Jaroslava
Podklady pro metodiku vzorkování rostlinných materiálů pro potřeby správních úřadů. Zpracování informací z ENGL pro potřeby ČIŽP. Shromažďování referenčních materiálů. Metodiky stanovení GMO. Svazek obsahuje sborník na téma Geneticky modifikované organismy - současnost, rozšíření a možné interakce s životním prostředím. Jednotlivé příspěvky ve sborníku: geneticky modifikované organismy - základní informace, Možnosti stanovení GMO, Křižitelnost plodin a planých druhů jako potenciál pro únik genetických modifikací, Šíření pylu v porostech lnu setého, Problematika GM hrachu v ČR, Perspektivy využití GMO, Problematika GM bramboru v ČR.
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PCR based assay for detection garden pea lec gene
Vráblík, Aleš ; Hodek, Jan ; Ovesná, Jaroslava
The method was developed and verified by staff of National reference laboratory for GMO identification and DNA fingerprinting suited for governmental control laboratories and private laboratories that analyze food and feed derived from various plant matrices. Method can be applied for quantification of GM pea in a sample, when reference gene is used as a standard. The method describe internal garden pea specific gene, lektin in this case, detection using PCR and real-time PCR. The general principle of the assay relies on lektin specific sequence presence that represents a species specific gene of garden pea. In many matrices DNA is damaged so amplification of short stretches of DNA is used. After amplification of target exploiting newly developer species specific gene primers PCR products are separated by electrophoresis in agarose gel. Resulting band is visualized by UV light and its position is compare with size standard. Alternatively, real-time PCR and ABI platform in combination with SYBR® Green can be used. Reaction parameters are described and specificity of reaction was verified. LOD as well as LOQ was defined.
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Methods of DNA extraction from the fresh papaya fruit and from the candied
Ovesná, Jaroslava ; Hodek, Jan
Aim the work is to provide a working procedure for extraction of amplifiable DNA form papaya fruits (Carica papaya), which are a poor source of DNA. Methods are optimised for DNA extraction from papaya fruits and stones and from candied papaya- these both were predicted as suitable commodities for an eventually screening of non-approved GM papaya in the market of Czech Republic. The methodology was prepared on basis of demand of reference laboratories of public service, which are involved in GMO analyses and also as a flow-work of Methodics for Detection of GM papaya 55-1 and 63-1 (Hodek, Ovesná, Pavlátová 2008) and scientific publication Detection of transgenic papaya lines: extraction protocol optimisation and verification of DNA quality by PCR assay (Ovesná, Hodek 2009).
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