Solo rna seq library preparation kit

RNA-seq libraries were built by the UPMC Children’s Hospital Health Sciences Sequencing Core following rRNA depletion. MNase-seq and ChIP-seq libraries were built prior to sample submission to the sequencing core. Libraries were prepared using the NEBNext Ultra II kit (New England Biolabs, Ipswitch, MA; DNA – E7645) and NEBNext Ultra sequencing indexes (NEB; E7335, E7500, E7710) according to manufacturer’s instructions. 4tU-seq libraries were built using a custom SoLo RNA-seq library preparation kit (TECAN, Redwood City, CA; 0516–32) with custom primers targeting S. cerevisiae rRNAs for depletion. All RNA and DNA libraries were quantified using Qubit and TapeStation and pooled for paired-end sequencing on an Illumina NextSeq 500 (UPMC Children’s Hospital Health Sciences Sequencing Core).

Libraries were prepared using the Tecan Ovation SoLo RNA-Seq library preparation kit as per the manufacturer’s instructions. Libraries were quantified using the Quant-iT PicoGreen dsDNA assay kit and read on a BioTek FLx800 fluorescence microplate reader. Libraries were pooled equal molar and the sample pool was quantified using Biorad ddPCR. Libraries were sequenced as stated below.