Naslov
Dizajniranje i obilježavanje sonde za humane gene 18S rDNA
(Design and labeling of the probe for the human 18S rDNA genes)

Autori
Chowdhury, Lily

Vrsta, podvrsta i kategorija rada
Ocjenski radovi, diplomski rad, diplomski

Fakultet
Prirodoslovno-matematički fakultet

Mjesto
Split

Datum
07.11

Godina
2014

Stranica
43

Mentor
Puizina, Jasna

Ključne riječi
primer design; fluorescent in situ hybridization; 18S rRNA genes; polymerase chain reaction
(primer design; fluorescent in sity hybridization; 18S rRNA genes; polymerase chain reaction)

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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