Electronic Journal of Biotechnology
versión On-line ISSN 0717-3458
ZHU, Houze et al. Improved isolation of good-quality total RNA from the optic stalk of Mud crab, Scylla paramamosain. Electron. J. Biotechnol. [online]. 2012, vol.15, n.2, pp. 5-5. ISSN 0717-3458.
An improved and efficient protocol was developed based on the TaKaRa RNAiso Plus Kit (Code: D9108A) for isolating good-quality total RNA from the optic stalk of mud crab, Scylla paramamosain. The protocol was based on the Trizol method with modifications. The carapace overlapping the optic stalk was retained with RNA in regular protocol. In order to remove the abundant deposition correlative with the carapace which makes the isolation of RNA particularly difficult, 5M potassium acetate solution (pH = 6.0) was added before the precipitation of RNA, and the temperature of RNA deposition was also decreased to -70ºC to ensure the stabilization of RNA. Good-quality total RNA from the optic stalk of S. paramamosain could be easily isolated with this modified protocol and three conventional methods were also employed to confirm the quality of RNA. This improved method would be helpful in facilitating molecular research of crabs involving RNA from the optic stalk.
Palabras clave : optic stalk; potassium acetate; RNA isolation; Scylla paramamosain.