Rapid isolation of double-stranded RNAs from entomopathogenic species of the fungus Paecilomyces using a commercial minicolumn system.
A method is described for rapid extraction of double-stranded RNAs from entomopathogenic fungi. Lyophilised and ground mycelium is incubated with 6 M guanidine thiocyanate, centrifuged, and the cleared lysate applied to a QIAGEN silica-based mini-spin column. Following washing with 70% isopropanol, bound nucleic acids are eluted under low salt conditions and treated with DNAse I prior to analysis by non-denaturing agarose gel electrophoresis.
Inglis
P W
PW
Centro National de Pesquisa de Recursos Genéticos e Biotecnologia (CENARGEN/EMBRAPA), Brasília, DF, Brazil. peter@cenargen.embrapa.br
Valadares-Inglis
M C
MC
D000818:Animals; D001203:Ascomycota / Q000737:chemistry*; D007313:Insecta / Q000382:microbiology; D012330:RNA, Double-Stranded / Q000302:isolation & purification*; D012331:RNA, Fungal / Q000302:isolation & purification*
D012330:RNA, Double-Stranded; D012331:RNA, Fungal