title: GSM4222316: HFD.20wk.3.technical_replicate.1; Mus musculus; RNA-Seq
library_construction_protocol: Isolated adipocytes were snap frozen in trizol were thawed on ice and then transferred to Fastprep matrix D lysing tubes (Mp Biomedicals) for homogenization using a Qialyzer instrument (Qiagen). Cells were homogenized with 2 cycles of 2 minutes at 20 1/s frequency, rotating tubes within holders (outside to inside and vice-versa) between rounds. RNA was extracted from the adipocyte homogenates using the RNeasy Lipid Tissue Mini kit and the Qiagen user-developed protocol titled Purification of total RNA from fatty tissues using the RNeasy® Lipid Tissue Mini Kit and MaXtract High Density (Qiagen). On-column DNA digest was performed for all samples according to manufacturer protocol (Qiagen). RNA was quantified using a Nanodrop 8000 spectrophotometer and quality was assessed using an Agilent 4100 Bioanalyzer (Agilent). cDNA libraries for RNA-seq analysis were prepared from total RNA (isolated as described above) according to Illumina TruSeq stranded mRNA sample prep LS protocol.
attributes.tag: GEO Accession
attributes.value: GSM4222316