NextSeq 500 paired end sequencing; Single cell transcriptional profile of third instar larval wing discs in Drosophila
Single cell transcriptional profile of third instar larval wing discs in Drosophila
Wandering third instar larvae were selected once outside the food  Flies were kept at 25 Â°C in vials containing standard cornmeal- agar medium. Third instar larva were collected, wing discs were dissected, pouch was manually removed using a microblade. Tissue was dissociated into single-cell suspension using a combination of Trypsin and Collagenase. Then  Drop-seq was performed on the single cell suspension. Libraries were made and sequenced. 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
Experimental Factor: RNA interference
mCherry knockdown by RNAi