文章基本信息

标题：Rapid RNA analysis of individual Caenorhabditis elegans
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作者：Kien Ly ; Suzanne J. Reid ; Russell G. Snell 等
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