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Extraction of RNA of high quality and yield from Aspergillus niger mycelia could appear difficult, owing to the highly active RNases released especially at the initial step of homogenising Aspergillus niger mycelia or at the final step when the RNA is solubilised and easily accessible to the RNAses. These two steps remain crucial for the successful extraction of RNA from A. niger mycelia. The total RNA being extracted might be partially or even totally degraded and the yield is limited. However, the inclusion of an RNase inhibitor (RNaseOUT) into these crucial RNA extraction steps solves the problem such that the RNase being released is inhibited by RNAseOUT and as such gives room for a good yield and high quality RNA which is of utmost importance when reverse transcription polymerase chain reaction (RT-PCR) is to be carried out or when gene expression profiles are to be investigated. Different methods of RNA extraction have been described (Cathala et al., 1983; David 2007; Loens et al., 2008), each of which did not address the peculiar problem encountered when extracting RNA form A. niger mycelia. Even methods of Salzman et al.(1999), Gehrig et al.(2000), and more recently Davis et al.(2006) that were specifically for extracting RNA form plant species did not directly address the peculiarity of the A. niger mycelia. This article therefore presents an extraction protocol developed for extracting RNA from A. niger mycelia with high success which is a modification of a standard operation procedure for RNA extraction using TRIZOL. Effectiveness of this protocol was tested by carrying out reverse transcription of the RNA followed by PCR …