title: Rapid RNA analysis of individual Caenorhabditis elegans.
abstract: Traditional 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 from Caenorhabditis elegans presents 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.
authors.affiliation: School of Biological Sciences, The University of Auckland, Auckland 1010, New Zealand.
authors.forename: Kien
authors.initials: K
authors.lastname: Ly
authors.affiliation: School of Biological Sciences, The University of Auckland, Auckland 1010, New Zealand.
authors.forename: Suzanne J
authors.initials: SJ
authors.lastname: Reid
authors.affiliation: School of Biological Sciences, The University of Auckland, Auckland 1010, New Zealand.
authors.forename: Russell G
authors.initials: RG
authors.lastname: Snell
keywords: C. elegans; RNA; RT-qPCR; Rapid single worm RT-qPCR