GSM7611261: Scale S3; Danio rerio; RNA-Seq
For procedures, zebrafish subjects were anesthetized with 0.02% tricaine methanesulfonate. Thymus, kidney marrow and scale samples were dissected and placed in 500 ul cell media (RPMI + 1% FBS + 1% Pen/Strep). Single-cell suspensions were prepped by dissociating tissues using a pestle and passed through 35 um filters. Various fluorescent populations were sorted from the lymphoid and precursor gates using a BD-FACSJazz Instrument (analysis performed in FlowJo softare). Following creation of single-cell emulsions, uniquely-identifiable 1st-strand template single-cell cDNA libraries were generated from each cell by emulsion PCR. 2nd-strand cDNA was generated and ligated to compatible Illumina adapters. Libraries were loaded onto single NovaSeq 6000 lanes and sequenced, using read lengths of 28 bp for the first read, 120 bp for the second read, and 8 base index reads.