GSM3523229: IL4 injected GFP(-) 24 hpi rep2; Danio rerio; RNA-Seq
Zebrafish brains from Tg(her4.1:EGFP) were dissected after PBS or IL4 injections at 24 hpi. Cells were dissociated with Miltenyi Neural Tissue Dissociation Kit as described (Kyritsis et al., 2012). Cells were sorted with BD Aria II FACS machine, RNA was isolated using Norgen Total RNA Purification Kit. Complete cDNA was synthesized from the obtained mRNA with SmartScribe reverse transcriptase using a universally tailed poly-dT primer and template switching oligo. This was followed by amplification of the purified cDNA with the Advantage 2 DNA Polymerase. After ultrasonic shearing of the amplified cDNA (Covaris S2) samples were subjected to standard Illumina fragment library preparation using the NEBnext chemistries (New England Biolabs). Libraries were purified using XP beads (Beckman Coulter), quantified by qPCR (KAPA Biosysytems) and subjected to Illumina 75bp single-end sequencing on the Illumina HiSeq2000 platform providing on average 31 million reads per sample.
GEO Accession
GSM3523229