GSM8566334: Ndufs2-KO-2; Mus musculus; RNA-Seq
Fresh gonadal white adipose tissue was minced in digestion buffer (2.5 units/ml collagenase D, 2.4 units/ml Dispase II, 10 mM CaCl2 in PBS) followed by incubation in a shaking water bath at 37C. The homogenate was then filtered through a 100 um cell strainer, followed by centrifugation at 600g for 5 minutes. The floating adipocyte fraction was isolated with a 1 mL pipette, washed with PBS, and then added to Trizol followed by RNA isolation using the Qiagen RNeasy mini kit. Libraries were constructed from 250 ng of RNA using the NEBNext RNA Ultra Directional library prep kit with poly(A) enrichment. Average library size and concentration were determined by Bioanalyzer and qPCR (NEBNext Library Quant kit)