title: GSM6380744: ABD adipose tissue of 2 apple-shaped and 2 pear-shaped subjects; Homo sapiens; RNA-Seq
library_construction_protocol: Cells were harvest at 80% confluency using trypsin. Selected cell were subjected to first-strand cDNA synthesis initiated by oligo dT primer (SMART-Seq ICELL8 CDS), followed by template switching with template switching oligo (SMART-Seq ICELL8 oligonucleotide) for 2nd strand cDNA synthesis, prior to unbiased amplification of full-length cDNA. Synthesized full-length cDNA was tagmented by TDE1 (Illumina Tagment DNA enzyme) and amplified using forward (i5) and reverse (i7) indexing primers. Each single cell/nucleus was indexed by a unique combination of 1 of 72 forward and 1 of 72 reverse indexing primers allowing for downstream identification. Collected cDNA was purified twice using a 1:1 proportion of AMPure XP beads (Beckman, A63880), further amplified according to manufacturer's instructions and purified again at a 1:1 proportion of AMPure XP beads. The resultant sequencing-ready cDNA librairy was assessed for concentration by fluorometer (Qubit) and quality by electrophoresis (Agilent Bioanalyzer high sensitivity DNA chips) respectively. Librairies were sequenced with Illumina HiSeq X