| Summary: | To survive in extremely cold environments, psychrophilic microorganisms produce exopolysaccharides (EPS), which are
carbohydrate polymers that constitute a substantial component of the extracellular polymers surrounding cells of these
microorganisms. EPS can interfere with RNA extraction and decrease the purity of the RNA extracted from EPS producing
microorganisms. In this work, six commercial RNA extraction kits and two published protocols for RNA extraction were
evaluated for total RNA extraction from the psychrophilic yeast, Glaciozyma antarctica. All the protocols were optimised to
obtain the highest quality of total RNA. The results show that all of the tested commercial kits and the tested conventional
methods yielded RNA from G. antarctica, albeit with varying quality. The protocol that utilises TRIzol® reagent was the
most effective method for isolating total RNA from G. antarctica of which this protocol resulted in the highest RNA yield
and purity compared to other methods. This method of RNA extraction produced RNA of sufficient quality for reverse
transcriptase PCR (RT-PCR) to detect the expression of the G. antarctica delta 9-fatty acid desaturase gene as well as for the
construction of a G. antarctica cDNA library.
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