Versuche zur Etablierung eines Verfahrens zur Isolierung von RNA aus Eberspermien
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vor 17 Jahren
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.
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.
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