title: GSM2108406: undifferentiated KD3 myoblast single-nucleus RNA-seq 4; Homo sapiens; RNA-Seq
library_construction_protocol: The cell membrane was ruptured with cold cell lysis buffer (10mM Tris-HCl, pH 7.4, 10mM NaCl, 3mM MgCl2, 0.005% IGEPAL CA-630) for 4-5 mins to release nucleus and the lysis efficiency was checked by Trypan blue (Lonza) staining. Single cells and nuclei were isolated using the Fluidigm C1 System. We used the largest integrated fluidic circuits (IFCs) of 17-25mm for KD3 cells. Single nuclei C1 runs were completed using the smallest IFC (5-10mm).  After capturing, the IFC was loaded with Clontech SMARTer kit lysis, RT, and PCR amplification reagents. Lysis, RT, and PCR amplification was completed overnight and harvested the following day. After harvesting, libraries were constructed using Illumina’s Nextera XT library prep kit per Fluidigm’s protocol. Constructed libraries were multiplexed and purified using AMPure beads.
attributes.tag: GEO Accession
attributes.value: GSM2108406