Home > Search Results: keywords:"syntéza DNA,GCPII,rakovina prostaty,qPCR,expresní vektor"
Search:
Search Tips :: Advanced Search
Search collections:
Sort by: Display results: Output format:
National Repository of Grey Literature 2 records found  Search took 0.00 seconds. 
Synthesis of 5'-UTR of the PSM-E variant of GCPII gene as a standard for RT-PCR
 Růžičková, Barbora ; Ingr, Marek (advisor) ; Martínek, Václav (referee)
PCR method has recently become a key element for the syntesis of genes. In this work two-step DNA synthesis based on PCR was used in order to obtain 5'-UTR of the PSM-E splicing variant of glutamate carboxypeptidase II (GCPII) which should serve as a standard for qPCR in real time. Another aim of this work was to clone this sequence into the plasmid PCRII-TOPO PSM-E for the purpose of protein expression in insect cells in the baculovirus expression system. In the first PCR the middle section of the 5'-UTR sequence was synthesized. This product was subsequently amplified in the second PCR using the first and last oligonucleotide. The product of the second PCR was ligated into the plasmid pUC19 and E. coli was transformed by tis construct. Sequence correctness was confirmed by sequencing analysis. Subcloning of the insert from pUC19 to pCRII-TOPO PSM-E was carried out and the sequence was checked by sequencing analysis again. The obtained construct was used as a standard in qPCR in real time in order to determine the concentration of the samples of mRNA from the prostate tissues of oncologic patients
Detailed record
Synthesis of 5'-UTR of the PSM-E variant of GCPII gene as a standard for RT-PCR
 Růžičková, Barbora ; Ingr, Marek (advisor) ; Martínek, Václav (referee)
PCR method has recently become a key element for the syntesis of genes. In this work two-step DNA synthesis based on PCR was used in order to obtain 5'-UTR of the PSM-E splicing variant of glutamate carboxypeptidase II (GCPII) which should serve as a standard for qPCR in real time. Another aim of this work was to clone this sequence into the plasmid PCRII-TOPO PSM-E for the purpose of protein expression in insect cells in the baculovirus expression system. In the first PCR the middle section of the 5'-UTR sequence was synthesized. This product was subsequently amplified in the second PCR using the first and last oligonucleotide. The product of the second PCR was ligated into the plasmid pUC19 and E. coli was transformed by tis construct. Sequence correctness was confirmed by sequencing analysis. Subcloning of the insert from pUC19 to pCRII-TOPO PSM-E was carried out and the sequence was checked by sequencing analysis again. The obtained construct was used as a standard in qPCR in real time in order to determine the concentration of the samples of mRNA from the prostate tissues of oncologic patients
Detailed record

Interested in being notified about new results for this query?
Subscribe to the RSS feed.
Digital Repository :: Search :: Submit :: Personalize :: Help
Powered by Invenio v1.1.2
Maintained by admin.invenio@techlib.cz
This site is also available in the following languages:
Česky  English