Illumina NovaSeq 6000 sequencing; scRNA-seq of lung stromal cells (CD45-CD31-CD326-) from C57BL/6J mice
scRNA-seq of lung stromal cells (CD45-CD31-CD326-) from C57BL/6J mice
Sample 1_p
Naïve C57BL/6J mice were euthanized, and lung was harvested and digested. Lung single cell suspension was prepared and stained with primary-fluorophore conjugated antibodies. Lung stromal cells (CD45- CD31- CD326-) were isolated by fluorescence-activated cell sorting. Cells capture, lysis and RNA extraction were performed using the 10x V3 Chromium technology. Around 12,000 cells were loaded in each lane of the 10x Chromium chip. Barcoding and library preparation were performed with the 10x Chromium system using version 3 chemistry, following the manufacturer's protocol (10x Genomics). cDNA was synthesized and amplified as per manufacturer's protocol and amplified to construct an Illumina sequencing library. cDNA and libraries were checked for quality on an Agilent 4200 TapeStation, quantified by KAPA qPCR.