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Versuche zur Etablierung eines Verfahrens zur Isolierung von RNA aus Eberspermien
Versuche zur Etablierung eines Verfahrens zur Isolierung von RNA aus Eberspermien
The aim of this work was to establish an optimal method for RNA isolation from the boar spermatozoa. This method should meet the following requirements: maximum amount of extracted RNA with high level of purity and avoidance of contamination by genomic DNA. These parameters were examined and judged by gel electrophoresis and quantitative real time PCR with cDNA- and DNA-specific primers. The TriZol and RNA-pure protocols both fulfil the requirements regarding quality and amount of isolated RNA. In contrast, the RNeasy method isolated RNA with genomic DNA contamination. The other four protocols allow isolation of “clean” RNA as well, but the quantity of RNA which could be recovered by these methods was less as compared to the TriZol and RNA-pure extraction procedures.
RNA isolation, boar spermatozoa, TriZol, RNA-Pure and SDS/DTT/Proteinase K, PRM2
Savchuk, Lika
2006
German
Universitätsbibliothek der Ludwig-Maximilians-Universität München
Savchuk, Lika (2006): Versuche zur Etablierung eines Verfahrens zur Isolierung von RNA aus Eberspermien. Dissertation, LMU München: Faculty of Veterinary Medicine
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Abstract

The aim of this work was to establish an optimal method for RNA isolation from the boar spermatozoa. This method should meet the following requirements: maximum amount of extracted RNA with high level of purity and avoidance of contamination by genomic DNA. These parameters were examined and judged by gel electrophoresis and quantitative real time PCR with cDNA- and DNA-specific primers. The TriZol and RNA-pure protocols both fulfil the requirements regarding quality and amount of isolated RNA. In contrast, the RNeasy method isolated RNA with genomic DNA contamination. The other four protocols allow isolation of “clean” RNA as well, but the quantity of RNA which could be recovered by these methods was less as compared to the TriZol and RNA-pure extraction procedures.