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Izolace RNA třešně
Vojáčková, Karolína ; Sedlák, Petr (vedoucí práce) ; Korecký, Jiří (oponent)
This bachelor thesis deals with the Isolaton RNA in sweet cherries. The turning point was the literature review on this topic. First chapters mentions the problemes of studying two main topics, which are RNA and a description of cherry (Prunus avium). Sweet cherry is a part of Noble hardwoods. Researches of sweet cherries can deal with an insufficient amount of quality planting material needed to breed new varieties. The biggest attention has given to the methods of RNA isolation, their comparison and analysis of already isolated RNA. The work describes five methods most commonly used for the isolation of RNA by phenol-chloroform, adsorption to silica, CTAB, TRIzol, magnetic separation and the using commercially marketed kits.
Phenol-chloroform method divides the lysate with a mixture of phenol, chloroform and isoamylalcohol in two phases, an upper aqueous, that includes the nucleic acids and lower organic.
Adsorption on silica using properties of nucleic acids bind to the surface of the silica in the presence of chaotropic salts.
CTAB method involves the use of extraction buffer with CTAB, which forms a precipitate with the nucleic acids. The pellet is then resuspended in DEPC-water.
TRIzol method uses a combination of guanidinium thiocyanate-phenol-chloroform. Like the phenol-chloroform method produces a precipitate, but treatment with guanidine thiocyanate, upper phase contained only RNA.
During the magnetic separation are the nucleic acids bound to the surface of the magnetized particles.
Subsequent genetic analysis that can detect and quantify the isolated RNA may be used in RT-PCR analysis, electrophoresis, and microarray hybridization methods northern blot and dot blot.
Bachelor thesis has written as a compilation literature review to inform the about mentioned topics. Any own research part is not here, because of the compilation character.
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