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| Description
| - Hsp proteins have been implicated in various cellular pathological events. We focused on expression of the Hsp70 protein family in the human hepatocyte HepG2 cell line and attempted to elaborate a method for its monitoring at the mRNA level using LightCyclerTM instrument (Roche). Cell lysates were prepared and purification of mRNA was done by either the magnetic separation method (Dynal) or using a High Pure RNA isolation kit (Roche). Corresponding cDNA was synthesized first. Six primer pairs for the Hsp70 family were examined. Among them, both primers designed in this laboratory and primers adopted from literary data were examined. Amplification reactions were effected in SYBR Green detection format. Purity and length of PCR products as well as those of the source mRNA were checked electrophoretically.As a result, an optimized primer pair giving a product of adequate degree of purity and expected chain length was selected.The method will be utilized for monitoring Hsp70 family mRNA levels in si
- Hsp proteins have been implicated in various cellular pathological events. We focused on expression of the Hsp70 protein family in the human hepatocyte HepG2 cell line and attempted to elaborate a method for its monitoring at the mRNA level using LightCyclerTM instrument (Roche). Cell lysates were prepared and purification of mRNA was done by either the magnetic separation method (Dynal) or using a High Pure RNA isolation kit (Roche). Corresponding cDNA was synthesized first. Six primer pairs for the Hsp70 family were examined. Among them, both primers designed in this laboratory and primers adopted from literary data were examined. Amplification reactions were effected in SYBR Green detection format. Purity and length of PCR products as well as those of the source mRNA were checked electrophoretically.As a result, an optimized primer pair giving a product of adequate degree of purity and expected chain length was selected.The method will be utilized for monitoring Hsp70 family mRNA levels in si (en)
- Hsp se účastní různých patologických dějů v buňkách. Zaměřili jsme se na expresi proteinů rodiny Hsp70 v buněčné liniilidských hepatocytů HepG2. Cílem bylo vypracovat metodu pro monitorování těchto proteinů na úrovni mRNA s použitím přístroje LighCycler (Roche).Z buněčných lysátů byly purifikovány RNA magnetickou separací (souprava Dynal) nebo High Pure RNA isolation kitem(Roche). Byla syntetizována odpovídající cDNA. Pro Hsp70 jsme testovali šest párů primerů, mezi nimi dva primery navržené naší laboratoří a primery převzaté z literatury. Amplifikační reakce probíhaly v detekčním formátu SYBR Green I. Čistota a délka PCR produktů a výchozí mRNA byly ověřeny elektroforézou. Vybrali jsme optimální pár primerů, poskytující produkt odpovídajícího stupně čistoty a očekávané délky řetězce. Metoda bude využita pro monitorování hladin mRNA rodiny Hsp70 ve studiích signálních kaskád HepG2 buněk, které v současnosti v naší laboratoři probíhají. (cs)
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| Title
| - Vývoj a optimalizace real-time PCR metody pro monitorování exprese proteinů rodiny Hsp70 v HepG2 buňkách (cs)
- DEVELOPMENT AND OPTIMIZATION OF A REAL-TIME PCR METHOD FOR MONITORING Hsp70 PROTEIN FAMILY EXPRESSION IN HepG2 CELLS
- DEVELOPMENT AND OPTIMIZATION OF A REAL-TIME PCR METHOD FOR MONITORING Hsp70 PROTEIN FAMILY EXPRESSION IN HepG2 CELLS (en)
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| skos:prefLabel
| - Vývoj a optimalizace real-time PCR metody pro monitorování exprese proteinů rodiny Hsp70 v HepG2 buňkách (cs)
- DEVELOPMENT AND OPTIMIZATION OF A REAL-TIME PCR METHOD FOR MONITORING Hsp70 PROTEIN FAMILY EXPRESSION IN HepG2 CELLS
- DEVELOPMENT AND OPTIMIZATION OF A REAL-TIME PCR METHOD FOR MONITORING Hsp70 PROTEIN FAMILY EXPRESSION IN HepG2 CELLS (en)
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| skos:notation
| - RIV/61989592:15110/04:00000838!RIV/2005/MSM/151105/N
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| http://linked.open.../vavai/riv/strany
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| http://linked.open...avai/riv/aktivita
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| http://linked.open...avai/riv/aktivity
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| http://linked.open...iv/cisloPeriodika
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| http://linked.open...vai/riv/dodaniDat
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| http://linked.open...aciTvurceVysledku
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| http://linked.open.../riv/druhVysledku
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| http://linked.open...iv/duvernostUdaju
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| http://linked.open...titaPredkladatele
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| http://linked.open...dnocenehoVysledku
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| http://linked.open...ai/riv/idVysledku
| - RIV/61989592:15110/04:00000838
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| http://linked.open...riv/jazykVysledku
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| http://linked.open.../riv/klicovaSlova
| - Hsp70;real-time PCR;HepG2 (en)
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| http://linked.open.../riv/klicoveSlovo
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| http://linked.open...odStatuVydavatele
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| http://linked.open...ontrolniKodProRIV
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| http://linked.open...i/riv/nazevZdroje
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| http://linked.open...in/vavai/riv/obor
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| http://linked.open...ichTvurcuVysledku
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| http://linked.open...cetTvurcuVysledku
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| http://linked.open...vavai/riv/projekt
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| http://linked.open...UplatneniVysledku
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| http://linked.open...v/svazekPeriodika
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| http://linked.open...iv/tvurceVysledku
| - Veselý, Jaroslav
- Uherková, Lenka
- Rypka, Miroslav
- Červenková, Kateřina
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| issn
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| number of pages
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| http://localhost/t...ganizacniJednotka
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