title: GSM9059204: All 3 neutrophil cultures (Figure 6) GEX; Mus musculus; RNA-Seq
library_name: GSM9059204
library_construction_protocol: For pooling purposes, bone marrow from control and AML mice were stained with TotalSeq-B hashtaging antibodies as per the protocol provided by BioLegend. Live singlets gated on host neutrophils were sorted in DPBS 0.04% BSA before loading onto 10x Genomics instrument according to the manufacturer's protocol. SImilarly, all three neutrophil culture conditions were pooled together along with hashtaging antibodies and processed as above. Gene expression and feature barcode libraries for single-cell RNA-seq were prepared using the Chromium Next GEM Single Cell 3' Gene Expression v3 with modifications for hashtag libraries. Quality control for amplified cDNA libraries and sequencing libraries was performed using a BioAnalyzer High Sensitivity DNA kit (Agilent). GEX and Cell Hashing libraries were sequenced on Illumina NextSeq 500 with a sequencing depth of at least 50,000 reads per cell for 3' GEX and 5,000 reads per cell for feature barcode.