A comparison of three methods for isolating RNA from mosquitoes.

The two most common protocols for RNA isolation, Chirgwin et al., 1979, and Chomczynski & Sacchi, 1987, were compared with a protocol using glass powder. The quality of the recovered RNA was tested using agarose gels and Northern hybridization. With the glass powder protocol, we consistently obtained intact RNA, whereas with the other protocols we often obtained degraded RNA. Using specific radioactive probes, we measured ribosomal RNA and trypsin mRNA recovered from fed adult Aedes aegypti females. Our protocol gave a 40- and a 3-fold increase in recovery when compared with the Chirgwin and Chomczynski protocol, respectively. The method using glass powder is simple, fast, reproducible and gives a high yield of intact RNA.