GSM9081192: Glomeruli, 6dpf, NFP-treated, 3; Danio rerio; RNA-Seq
For total RNA, whole larvae samples were processed using Tri-Reagent (Sigma-Aldrich) according to manufacturer's instructions, followed by DNA-digestion and purification steps using the RNase-Free DNase I Kit and RNA Clean-Up and Concentration kit according to manufacturer's instructions (both Norgen Biotek Corp., Thorold, ON, Canada). The quality of the extracted RNA was assessed using the Bioanalyzer 2100 (Agilent, Santa Clara, CA, USA).  All samples exhibited a mean RNA integrity score (RIN) of 10. Total RNA of isolated glomeruli was derived by combination of Tri-Reagent (Sigma Aldrich) and Norgen Single Cell RNA Purification kit (Norgen Biotek Corp., Thorold, ON, Canada) according to manufacturer´s instructions with an elution volume of 10µl. The quality of the extracted RNA was assessed using the Bioanalyzer 2100 (Agilent, Santa Clara, CA, USA).  All samples exhibited a mean RNA integrity score (RIN) of 7.4 ± 0.3 SD. PolyA library was prepared based on 500ng total RNA per sample using the TruSeq stranded mRNA kit (Illumina, San Diego, CA, USA).  Subsequently, for sequencing 200pM of the final library were loaded on the NovaSeq 6000 System, (2 x 100 bp pair-end reads). A sequencing depth of ~ 200 Mio Cluster per sample was obtained.