1.6. Expression of fungal genes

Studies investigating host-pathogen interactions have taken a new dimension utilizing mRNA quantification (qRT-PCR). Here we describe qRT-PCR approach used for identification of A. apis mating type idiomorphs and quantification of A. apis transcripts in culture and in host tissue (Aronstein et al., 2007). Total RNA can be isolated using standard kits, TRIzol (Invitrogen; Carlsbad, California) and RNeasyR Mini Kit (Qiagen; Valencia CA). These reagents have been used successfully to make cDNA from mycelia or honey bee larvae (Aronstein et al., 2007; 2010), for RNA extraction method, see the BEEBOOK paper on molecular methods (Evans et al., 2013)