GSM7823339: SH005-GEX; Homo sapiens; RNA-Seq
iMG were incubated with TruStain FcX blocking reagent (BioLegend, Cat. No. 422301). Hash tagging antibodies were added, and cells were incubated for additional 30 minutes at 4C and washed.  Library preparation was conducted using 10x Chromium Next GEM Single Cell 3' Reagent Kits v3.1 with Feature Barcode technology. Library was constructed following manufacturer's portocol. Briefly, reverse transcription reagents, barcoded gel beads, and partitioning oil were mixed to generate GEM. After the reverse transcription, GEMs were broken, and Feature Barcode cDNA amplification occurred. Amplified cDNA was separated by SPRI size selection into mRNA and HTO fractions. Sequencing libraries were generated and analyzed using TapeStation D5000 and Qubit HS DNA quantification kit. Sequencing was performed on a NovaSeq 6000 with S4 flow cell (Illumina, San Diego, CA) using paired-end, dual-index sequencing as: 28 cycles for read 1, 10 cycles for i7 index, 10 cycles for i5 index, and 90 cycles for read 2.