title: GSM6675073: A, ZA4, scRNAseq [A21]; Homo sapiens; RNA-Seq
library_construction_protocol: All samples were incubated with 2 μL of 10× Reaction Buffer (SMART-Seq v4 Ultra-Low Input RNA kit for Sequencing, (Takara Bio, USA) and preserved at −80°C until preparation analysis. Total RNA was extracted using a SMART-Seq V4 Ultra-Low Input RNA Sequencing kit (Clontech, CA, USA) that uses oligo(dT) priming. cDNA was synthesized with 3′SMART-Seq CDS primer II (Clontech, CA, USA) and PCR amplified (17 cycles) from 10 pg of RNA following manufacturer's instructions. Briefly, cDNA was purified using AMPure XP magnetic beads (Illumina, USA). After confirming cDNA integrity on a 2100 Bioanalyzer (Agilent Technologies, USA), a total of 50 ng of genomic DNA were fragmented with the TruSight One Sequencing Panel Library (Illumina, USA) according to manufacturer's instructions. RNA-seq libraries were constructed using the same kit. Then, a RNA pool was generated using equal amounts of RNA (5 nM) per individual sample. Barcodes and adapters were included in the library to sequence and identify all samples.