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  • 标题:Rapid RNA analysis of individual Caenorhabditis elegans
  • 本地全文:下载
  • 作者:Kien Ly ; Suzanne J. Reid ; Russell G. Snell
  • 期刊名称:MethodsX
  • 印刷版ISSN:2215-0161
  • 电子版ISSN:2215-0161
  • 出版年度:2015
  • 卷号:2
  • 页码:59-63
  • DOI:10.1016/j.mex.2015.02.002
  • 语种:English
  • 出版社:Elsevier
  • 摘要:Graphical abstractDisplay OmittedAbstractTraditional RNA extraction methods rely on the use of hazardous chemicals such as phenol, chloroform, guanidinium thiocyanate to disrupt cells and inactivate RNAse simultaneously. RNA isolation fromCaenorhabditis eleganspresents another challenge due to its tough cuticle, therefore several repeated freeze–thaw cycles may be needed to disrupt the cuticle before the cell contents are released. In addition, a large number of animals are required for successful RNA isolation. To overcome these issues, we have developed a simple and efficient method using proteinase K and a brief heat treatment to release RNA of quality suitable for quantitative PCR analysis.The benefits of the method are:•Faster and safer compared to conventional RNA extraction methods•Released RNA can be used directly for cDNA synthesis without purification•As little as a single worm is sufficient
  • 关键词:C. elegans;RNA;RT-qPCR
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