title: GSM8143733: HFD, large nuclei, replicate2; Mus musculus; RNA-Seq
library_name: GSM8143733
library_construction_protocol: Nuclei were isolated from frozen inguinal (iWAT) white adipose tissues by pulverizing the tissues in liquid nitrogen and homogenizing in chilled Douncers. Next, the nucleus suspension was subject to nucleus sorting to remove debris and multiplets. RNA was extracted using TRIzol and used for constructing sequencing libraries. The extracted RNA (100ng) was first purified using the TURBO DNase Kit and processed by NEBNext rRNA Depletion Kit for removing ribosomal RNA. Then, RNA was converted to cDNA using Maxima Reverse Transcriptas and NEBNext mRNA Second Strand Synthesis Kit following manufacturer's protocols. After purification of cDNA based on size, sequencing libraries were generated using Nextera XT DNA Library Preparation Kit and subsequent PCR amplification.