title: GSM9240161: HFD2; Mus musculus; RNA-Seq
library_construction_protocol: The eWAT were isolated, minced, and digested in collagenase II. The digest was centrifuged, and the floating adipocytes were subsequently discarded. The SVF cells were filtered through a 40 μm cell strainer, centrifuged, and then re-suspended. ScRNA-seq libraries were generated using the 10x Genomics Chromium Single Cell 3' Library, following the manufacturer's instructions. Briefly, GCs were resuspended in the master mikx andloaded together with partitioning oil and gel beads into the chip to generate the gelbead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every singleGEM was retrotranscripted to cDNA, which contains an llumina R1 primer sequence.Unigue Molecular ldentifier (UMl) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiatedsized fragments were selected with SPRlselect reagent for subsequent libraryconstruction. During the library construction llumina R2 primer sequence, paired-endconstructs with P5 and P7 sequences and a sample index were added.