NextSeq 500 sequencing; Single-cell RNA-seq profiling of Drosphila melanogaster Rbf mutant third intsar eye discs
Single-cell RNA-seq profiling of Drosphila melanogaster Rbf mutant third intsar eye discs
Wandering third instar larvae were selected once outside the food. Rbf mutants were generated by P element insertion in the 5' untranslated region of Rbf which results in a hypomorphic mutation. The eye imaginal discs were separated from the antenna and brain. The tissues were dissociated using trypsin and collagenase Following dissociation, the single cells suspension was washed in PBS and 0.01% BSA as suggested in the Drop-seq protocol Cell were lysed in the droplets generated in Drop-seq. RNA was captured on oligo dT beads Library construction was performed as written in the Drop-seq protocol. 1) RNA was reverse transcribed. 2) cDNA was PCR amplified. 3) tagmentation was done using Nextera XT kit