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  • 标题:Inappropriateness of RNAlater to preserve Caenorhabditis elegans for RNA extraction
  • 本地全文:下载
  • 作者:Leming Jiang ; Linyan Li ; Ping Kang
  • 期刊名称:MethodsX
  • 印刷版ISSN:2215-0161
  • 电子版ISSN:2215-0161
  • 出版年度:2019
  • 卷号:6
  • 页码:2460-2467
  • DOI:10.1016/j.mex.2019.10.015
  • 语种:English
  • 出版社:Elsevier
  • 摘要:Graphical abstractDisplay OmittedAbstractCaenorhabditis elegansis 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 ofC. elegansfor gene expression studies. To address this issue, the present study has compared different conditions to preserveC. elegansfor 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 ofC. eleganswith 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 fromC. eleganswith variable worm numbers from 20 to 200.•RNAlater was inappropriate for preservingC. elegansfor effective RNA extraction.•Proteinase K was verified for lysing a limited number ofC. elegansfor RNA extraction.
  • 关键词:Caenorhabditis elegans;RNA extraction;RNAlater;Storage conditions;Proteinase K
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