GSM8032343: S28,Pilot experiment, scRNASeq; Homo sapiens; RNA-Seq
MCF-7 cells were washed twice with PBS, then placed into equilibrated Hank's balanced salt solution (HBSS) (Thermo Fisher Scientific, Waltham MA, USA) and maintained at 37°C during hyperspectral imaging. Subsequently, imaged cells were captured by CellCelector to a 384-well PCR plate with lysis buffer. Library was prepared following Smart-seq2 protocol. Briefly, cells were picked into 384 well lysis plates followed by reverse transcription, template switching and 26 cycles of PCR to generate and amplify the cDNA. MANTIS automated liquid handler (Tecan, Zürich, Switzerland) was used during the process. cDNA was normalised to 0.1 - 1ng/µl according to the qualification by Quant-iT™ PicoGreen™ dsDNA Assay Kit (Thermo Fisher Scientific, Waltham MA, USA) via automated liquid handling robot Mosquito (SPT Labtech, Melbourn, Cambridgeshire, UK). Nextera XT kit (Illumina, San Diego, CA, USA) with 14 cycles of amplification was used for tagmenting MCF-7 cells. Purification was conducted to remove primer by Agencourt Ampure XP (Beckman Coulter, Brea CA, USA) magnetic beads at a ratio of 0.7x for two times. Libraries were quantified using Bioanalyzer 2100 (Agilent Technologies, Waldbronn, Germany).