Inappropriateness of RNAlater to preserve Caenorhabditis elegans for RNA extraction
Citation
Jiang, L., Li, L., Kang, P., Yu, H., Nie, S.P., Xie, M.Y., Gong, J. (2019). Inappropriateness of RNAlater to preserve Caenorhabditis elegans for RNA extraction. MethodsX, [online] 6 2460-2467. http://dx.doi.org/10.1016/j.mex.2019.10.015
Plain language summary
Caenorhabditis elegans is a well-established laboratory animal model and has been widely used in biological research. However, it is still a challenge to obtain a good amount of quality RNA from a limited number of C. elegans for gene expression studies. To address this issue, the present study has compared different conditions to preserve C. elegans for RNA extraction after the failure of an initial effort to use the worms preserved in RNAlater. The effects of different concentrations of proteinase K, different worm life stages and numbers on RNA extraction were also investigated. A good amount of quality RNA was achieved under the following conditions: 1) adult worms that were either freshly prepared or quickly frozen in liquid nitrogen followed by storage at -80oC; 2) disruption of C. elegans with proteinase K (1 mg/mL, 65oC for 10 min) prior to extraction. The proposed method provides a stable, rapid, and effective means to extract RNA from C. elegans with variable worm numbers from 20 to 200.
Abstract
Caenorhabditis elegans is a well-established laboratory animal model and has been widely used in biological research. However, it is still a challenge to obtain a good amount of quality RNA from a limited number of C. elegans for gene expression studies. To address this issue, the present study has compared different conditions to preserve C. elegans for RNA extraction after the failure of an initial effort to use RNAlater-preserved worms for RNA extraction. The effects of different concentrations of proteinase K, different worm life stages, and different worm numbers on RNA extraction were also investigated. The best results were achieved under the following conditions: 1) adult worms that were either freshly prepared or quickly frozen in liquid nitrogen followed by storage at −80 °C; 2) disruption of C. elegans with proteinase K (1 mg/mL) in a lysis buffer (65 °C for 10 min) prior to extraction with Trizol agent. This method can provide a stable, rapid, and effective means to extract RNA from C. elegans with variable worm numbers from 20 to 200. • RNAlater was inappropriate for preserving C. elegans for effective RNA extraction. • Proteinase K was verified for lysing a limited number of C. elegans for RNA extraction.