Single cell Aplysia californica Neuronal Transcriptome
Single cell Neuronal Transcriptome form Aplysia 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.
Aplysia californica is one of key models to study cellular bases of behavior and plasticity. Its central nervous system composed of ~10,000 neurons most of them are unique and individually identifiable. Here, we performed transcriptome analysis of these neurons using pyrosequencing approach. 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 several thousands of neuronal transcripts (many of them are unique) including a diversity of receptors and transcription factors essential for the establishing of cell identity and plasticity.