Adapting the Smart-seq2 Protocol for Robust Single Worm RNA-seq

Author(s): Serra, Lorrayne; Chang, Dennis Z; Macchietto, Marissa; Williams, Katherine; Murad, Rabi; Lu, Dihong; Dillman, Adler R; Mortazavi, Ali | Abstract: Most nematodes are small worms that lack enough RNA for regular RNA-seq protocols without pooling hundred to thousand of individuals. We have adapted the Smart-seq2 protocol in order to sequence the transcriptome of an individual worm. While developed for individual Steinernema carpocapsae and Caenorhabditis elegans larvae as well as embryos, the protocol should be adaptable for other nematode species and small invertebrates. In addition, we describe how to analyze the RNA-seq results using the Galaxy online environment. We expect that this method will be useful for the studying gene expression variances of individual nematodes in wild type and mutant backgrounds.