title: GSM7505562: MTM1_set_2_S12; Danio rerio; RNA-Seq
library_construction_protocol: Total mRNAs were extracted from pools of dissected livers from 6 dpf abcb11b mutants and WT siblings, 20 livers per replicate and 3 replicates per genotype, by using Arcturus PicoPure RNA isolation kit (Applied Biosystems, KIT0204). For the 40 livers per replicate (4-5 replicates per group) were dissected under a brightfield microscope at 7dpf and then pooled prior to RNA extraction using the Qiagen RNAeasy Micro kit. Animals in the overfed group were provided with six times the normal amount of food at each feeding from day 5 to 7 while EtOH treated animals were exposed to 2% ethanol for 32 hours prior to sample collection. The presence of hepatic steatosis was confirmed in both groups. WT and abcb11b mutants samples were submitted to Novogene Co for library preparation and RNA-sequencing using the vendor's standard protocol.  The 40 samples were  submitted to the Centre for Applied Genomics (TCAG, Toronto, Canada). SMART-Seq v4 Ultra Low Input RNA kit was used for the cDNA conversion. Library preparation followed the protocol of Nextera XT library prep kit. Libraries were quality controlled (QC)ed (run on the Bioanalyzer DNA High Sensitivity chip to check for size and quantified by qPCR using Kapa Library Quantification Illumina/ABI Prism Kit protocol). All libraries were pooled and sequenced on 2 lanes on the NovaSeq SP flowcell paired-end 2x100bp.