title: GSM7115640: 468, Day0, replicate2, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA from cell lines were extracted using Qiagen RNeasy Mini kit (Qiagen, Hilden, Germany) following the manufacturer's instructions. A total of 1 μg of total RNA from each sample was used to obtain double strand cDNA with the QuantiTect Reverse Transcription Kit (Qiagen). MDAMB468 barcoded and DTP cells were trypsinized into single-cell suspensions and processed using a Chromium Single Cell Gene Expression 3′ Library and Gel Bead kit v3.1 following the manufacturer's instructions (10x Genomics). Estimated 1 x 104 cells were counted and checked for viability using a LUNA-II™ Automated Cell Counter (Logos Biosystems) and then injected into microfluidic chips with the average recovery rate of 5 x 103 cells to form single-cell Gel Beads-in-Emulsion (GEMs) in the 10x Chromium instrument. Reverse transcription was carried out on the GEMs, and reverse-transcribed samples were purified and amplified. The quality of the amplified cDNA was quantified with the BioAnalyzer High Sensitivity DNA kit D5000 (Agilent Technologies) and its concentration was measured using the Qubit Fluorometer. Finally, amplified cDNAs were fragmented, and adapter and sample indexes or custom primer to selectively amplify barcodes, were incorporated into the libraries which were compatible with Illumine next-generation sequencing. The size profiles and the quality of the obtained libraries were profiled using BioAnalyzer High Sensitivity DNA kit D1000. NovaSeq 6000 SP 100 cycles flow cell was used to sequence the libraries.