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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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Methodics of molecular detection of Barley yellow dwarf virus in vectors by RT-PCR
Jarošová, Jana ; Jaňourová, Blanka ; Kumar, Jiban
Barley yellow dwarf is one of the most important viral diseases of cereals in the world. It can cause significant yield losses in most cereal species, including wheat, barley, rice, corn, oat and rye. In this methodology we provide assays for reverse transcription - polymerase chain reaction (RT-PCR) for sensitive detection of barley yellow dwarf virus directly in the vectors, in aphids. This way it is possible to evaluate infectivity of vectors and from that conclude subsequent measures. The methodology is meant to serve the employees of State Phytosanitary Administration.
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Technical Means of Drug Control in Traffic.
ŠPALEK, Dušan
ABSTRACT The topic of the diploma thesis bears the title Possibilities of using technical means of narcotics checks in road traffic. The thesis endeavours to describe, compare and evaluate searching methods that are used in road checks for habit-forming drugs by public officials. The theoretical part is focused on the description of individual groups of habit-forming drugs and their effects on the human organism. Further, the theoretical part contains an overview of the most frequently used analytic methods. The practical part deals with the questions of detection aids used by the traffic police in the Czech Republic. Based on the methodology used, which included research in domestic and foreign sources and a structured interview, all the information obtained was thoroughly evaluated and it illustrates the substance of the topic. This diploma thesis is structured in such a way that enables its use by police units and other organisations that take part in fighting the use of habit-forming drugs. The thesis can be also a contribution for tuition as study material.
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