title: GSM8686089: HAP1_SC_3; Homo sapiens; RNA-Seq
library_name: GSM8686089
library_construction_protocol: Total RNA was extracted from biological replicates of 1, 10, 100 and 1000 sorted HAP1 cells. For each sample, cells were sorted with single cell precision using BD FACSAria IIu into a 60 µl mixture containing 1 part PBS and 2 parts NAxtra LYSIS BUFFER (Lybe Scientific) and combined with a 80 µl mixture of 4 µl NAxtra MAGNETIC BEADS (Lybe Scientific) in isopropanol. Following mixing and magnetization, the beads were washed on magnet with 150 µl absolute ethanol, dried for 2 min and resuspended in 15 µl nuclease-free water. Samples were DNase-treated by 10 min incubation with 0.4 µl RNase-Free DNase (QIAGEN), 2 µl Buffer RDD (QIAGEN) and nuclease-free water to a final volume of 20 µl. Nuclease-treated samples were combined with 40 µl NAxtra LYSIS BUFFER (Lybe Scientific), followed by 80 µl isopropanol. Following mixing and magnetization, the beads were washed on magnet, once with 150 µl isopropanol, and twice with 150 µl 80% ethanol, then dried for 2 min and resuspended in 8 µl nuclease-free water. Following mixing and magnetization, eluates were collected and stored at -80°C. The SMART-Seq® Stranded Kit (Takara) was used to generate dual index libraries according to manufacturer's instructions.