scRNA-seq of Homo sapiens: acute COVID, visit 5
Single-cell suspensions were washed twice with PBS with 0.04% BSA (Sigma-Aldrich), cells were counted and 30 000 cell input was used for further analysis with MGI DNBelab C Series High-throughput Single-cell RNA Library Preparation Set V2.0. In short, single cells were combined with magnetic beads and encapsulated in oil droplets using the DNBelab C4 microfluidic device to allow for mRNA hybridization. Magnetic beads were collected, mRNA from single cells was reverse transcribed and libraries were prepared according to the protocol.Resulting cDNA product fragment size was distributed between 1149bp to 1492bp. cDNA and Oligo libraries had fragment sizes between 428bp to 515bp and 181bp to 190bp accordingly. Quality control of fragment size and concentration was performed using Agilent 2100 Bioanalyzer System and Qubit dsDNA HS Assay kit for Qubit 2.0 Fluorometer (Thermo Fisher Scientific).