GSM7696905: Brain_Ipsi_Stroke_3day_Cells; Mus musculus; RNA-Seq
Cells were extracted from brains using a no-enzyme cold processing approach in Hank's balanced salt solution to avoid activation of immune cells due to warm enzyme processing. Single cell 3' gene expression (mRNA) and cell surface hashtag-oligonucleotide (HTO) libraries were prepared according to the Chromium Single Cell 3ʹ with Feature Barcoding technology for Cell Surface Protein Reagent Kit v3 protocol (10X Genomics) with modifications for incorporation of TotalSeq™-A HTO antibodies (clones M1/42; 30-F11, BioLegend). Briefly, generation of gel beads in emulsion (GEM) partitioned with single cells, reverse transcription to generate cDNA and HTO-barcoded DNA, and DNA amplification were performed according to manufacturer instructions with the addition of HTO primers during DNA amplification. Amplified cDNA and HTO-derived DNA were separated by size selection, fragmented and libraries constructed.