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National Repository of Grey Literature 1 records found  Search took 0.00 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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