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Design and labeling of the probe for the human 18S rDNA genes (CROSBI ID 392511)

Ocjenski rad | diplomski rad

Chowdhury, Lily Design and labeling of the probe for the human 18S rDNA genes / Puizina, Jasna (mentor); Split, Prirodoslovno-matematički fakultet u Splitu, . 2014

Podaci o odgovornosti

Chowdhury, Lily

Puizina, Jasna

engleski

Design and labeling of the probe for the human 18S rDNA genes

The aim of this work was to make a probe for human 18S rDNA genes, which could be further used for the physical mapping of 18S rDNA genes in the human karyotype by application of fluorescent in situ hybridization technique (FISH) for both teaching and research purposes. To this end, bioinformatics tools were used for the design and testing of two primers (20 nucleotide long single-stranded DNA molecules). According to the obtained sequences, the two primers, specific for the human 18S rDNA were synthesized and purchased from the specialized company. Those primers were used in polymerase chain reaction (PCR) to yield a specific PCR product, approx. 600 bp long, a partial human 18S rDNA. The PCR product was cloned in a plasmid and transformed into bacteria E. coli. Five clones were isolated and sequenced, and in three of them a specific PCR product has been confirmed. One of the clones has been labeled by the application of nick-translation procedure using the modified nucleotide, digoxigenin-dUTP. Thus, the main goal of this work, design and labeling of human 18S rDNA probes has been fully accomplished.

primer design; fluorescent in sity hybridization; 18S rRNA genes; polymerase chain reaction

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Podaci o izdanju

43

07.11.2014.

obranjeno

Podaci o ustanovi koja je dodijelila akademski stupanj

Prirodoslovno-matematički fakultet u Splitu

Split

Povezanost rada

Biologija