GSM8862540: AV13_02_Young_biological replicate 2; Mus musculus; RNA-Seq
RNA was extracted using Qiagen RNA extraction kit (Qiagen, Cat no. 74104) as per manual instructions. As for mammary fat pads, tissues were chopped into small pieces and digested in buffer using tungsten carbide beads (Cat no. 69997, Qiagen) and tissue lyser for 30s. Lysed tissues were immediately processed as per manual's instructions (Cat no. 74014, Qiagen). 1mg of purified RNA was sent for RNA sequencing analysis. Total RNASeq libraries were prepared using the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (New England Biolabs, E7760) and the NEBNext® rRNA Depletion Kit v2 (Human/Mouse/Rat) (New England Biolabs, E7400). An input of 1000ng of total RNA was used with 9 cycles of amplification.