Neuronal Transcriptome of Pleurobranchaea californica
Neuronal Transcriptome of Pleurobranchaea californica
We developed a reduced representation sequencing method that can detect low abundant transcripts. Our method greatly reduces the amount of sequencing with more accurate quantification. Basically, sequential addition of adaptors paired with controlled digestion of cDNA generates a one transcript on read type of analysis. Combined 5’ with 3’ targeted libraries produces greater coverage of individual transcripts and high quality assembly.
The Pacific sea slug, Pleurobranchaea californica is an important model to study cellular bases of behavior. However, nothing is known about the molecular organization of this nervous system. Here, using pyrosequencing approach we performed analysis of transcripts expressed in the central nervous system of Pleurobranchaea. Specifically, we developed a reduced representation sequencing method that can detect low abundant transcripts. Our method greatly reduces the amount of sequencing with more accurate quantification. Basically, sequential addition of adaptors paired with controlled digestion of cDNA generates a one transcript on read type of analysis. Combined 5’ with 3’ targeted libraries produces greater coverage of individual transcripts and high quality assembly.   Our initial analysis allowed us to identify more than 15,000 of neuronal transcripts including a diversity of novel neuropeptides, receptors, ionic channels, and other components of signal transduction pathways essential for future analysis of molecular bases of neuronal functions in this model organism.