Illumina HiSeq 4000 paired end sequencing; RNAseq analysis of 3T3-L1 adipocytes cultured in high and low media volumes
3T3-L1 fibroblasts were washed 3x in PBS and lysed in 350 µL RLT buffer. Lysed samples were collected and stored at -80°C until further analysis 3T3-L1 adipocytes were seeded and differentiated in 12-well plates. RNA extractions were performed using the RNeasy Mini Kit (Qiagen 1152 #74104) 1 μg of total RNA was quality checked (RIN >7) using an Agilent Bioanalyser 2100 system and then used to construct barcoded sequencing libraries with Illumina's TruSeq Stranded mRNA Library Prep Kit following manufacturer's instructions