6996m
RNA was prepared from frozen samples using Zymo Research Quick RNA MIcro Prep Kit # R1050 and cleaned using Zymo Research RNA Clean and Concentrator #R1015. Samples were analyzed on an Agilent QC 2100 Bioanalyzer to determine quality prior to library preparation. cDNA was prepared using the Nugen Ovation RNA Sequencing System V2, #7102, and cleaned using the QIAGEN QIAquick PCR purification kit, #28104, as recommended in the protocol, and analyzed on the Agilent QC Bioanalyzer. If needed, samples were size-selected using Zymo Research Select-A-Size DNA Clean and Concentrator #D4080 prior to barcoding. The final library was prepared using NEB NEBNext Ultra II DNA Library Prep Kit #27645 and barcoded using NEBNext Multiplex Oligos for Illumina #E6609S. All magnetic bead purification was accomplished using BullDogBio CleanNGS RNA and DNA Spri Beads #CNGS005. Samples were then analyzed using the Agilent QC 2100 Bioanalyzer to determine the concentration of each sample prior to sequencing.