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National Repository of Grey Literature 2 records found  Search took 0.01 seconds. 
The development of an HPLC method for monitoring the preparation of oligodeoxynucleotide probes
 Šemlej, Tomáš ; Kučera, Radim (advisor) ; Pilařová, Pavla (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Mgr. Tomáš Šemlej Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Title of rigorosum thesis: The development of an HPLC method for monitoring the preparation of oligodeoxynucleotide probes An artificially prepared oligodeoxynucleotide sequence with the attached label is referred to as a molecular probe. Molecular probes are mainly used to quantify multiplication in RT-PCR and identify mutations in PCR analyses. There are typically two different types of probes - mono-labeled molecular probes and double-labeled molecular probes. The mono-labeled molecular probes are constructed from fluorophore- labeled oligodeoxynucleotide sequence or quencher-labeled oligodeoxynucleotide sequence. Whereas double-labeled molecular probes consist of an oligodeoxynucleotide sequence with a fluorophore attached at one end together with a quencher attached at the other end of the chain. In this work, we first paid attention to the separation of double- labeled oligodeoxynucleotides from mono-labeled oligodeoxynucleotides. Several types of stationary phases were tested and based on that the column Clarity® Oligo-RP™ was chosen to separate a mixture of labeled oligodeoxynucleotides. This...
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Preparation of fluorescent azaphthalocyanines for labeling of oligonucleotide probes I.
 Beranová, Michaela ; Miletín, Miroslav (advisor) ; Kučerová, Marta (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical analysis Candidate Michaela Beranová Supervisor doc. PharmDr. Miroslav Miletín, Ph.D. Title of thesis Preparation of fluorescent azaphthalocyanines for labeling of oligonucleotide probes I. Azaphthalocyanines (AzaPc) are macrocyclic compounds with large system of conjugated bonds and are therefore used as dyes and pigments. Their potential as diagnostic and therapeutic means is also under intensive examination. The aim of this thesis is a synthesis of precursors for cyclotetramerization of asymmetric fluorescent tetrapyrazinoporphyrazines (TPyzPz) via solid phase method bearing two various moieties, one suitable for binding to solid phase and the other intended for binding of the final TPyzPz to oligonucleotides. At the beginning of the thesis, basic terms related to AzaPc are defined. Also the main principles of synthesis of TPyzPz as well as possible modifications of their final structure are discussed. An important part of the thesis is the description of the typical properties of TPyzPz, the influence of the structure on the individual parameters and the fields of their use. Afterwards the preparation of each precursor with two different substituents for cyclotetramerization is...
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