RNAseq MEFs Ube3a+/+ S1_L3
Ube3a-/- and Ube3a+/+ cultured for 5 passages as mentioned above were used for RNA sequencing. After trypsinization cells were collected and total RNA was isolated using the RNeasy Lipid Tissue Mini Kit, Cat No: 74804 (QIAGEN) according to the manufacturer instructions. Isolated RNA concentration and quality were determined by Qubit quantitation assay using Qubit 2.0 fluorometer (Invitrogen- life technologies, USA) and Agilent 4200 TapeStation System (Agilent Technologies, USA). Samples were prepared for Illumina sequencing using NEBs Ultra RNA Library Prep Kit for Illumina (NEB#7530) (BioLabs Inc, MA, USA) according to the manufacturer protocol.  Libraries were sequenced with a 2 X 150 bp PE run on Illumina HiSeq 2500 using a V3 flow cell.