GSM5829842: Amacrine cells/Horizontal cells rep4; Danio rerio; RNA-Seq
Retinal cell types were sorted from dissociated zebrafish retinas at 42 hpf using the triple transgenic line Tg(ath5:gap-RFP), Tg(crx:gap-CFP), Tg(ptf1a:gal4,UAS:gap-YFP). For each of the 5 replicates, 25 retinas were pooled.   During dissection, retinas were maintained at 4°C in a volume of 100 µl in PBS. Prior to dissociation, the volume was adjusted to 500 µl. Manual dissociation was performed immediately before FACS using BSA-coated glass pipettes.   500 cells per sample were FACS sorted directly into extraction buffer. Superscript II was used for reverse transcription and cDNA was amplified using Kapa HiFi HotStart Readymix (Peqlab). Purification of the amplified cDNA was performed using Ampure XP Beads (Beckman Coulter). Libraries were prepared accoridng to SmartSeq-2 flex Illumina protocol. Illumina sequencing was performed on a Nextseq500 with a sample sequencing depth of at least 30 mio reads.