GSM6257311: Basophils from Nb infected mice (hashtag library); Mus musculus; RNA-Seq
Single-cell suspensions were prepared from the bone marrow, spleen and infected skin and subjected to positive selection of CD49b-positive cells by using biotinylated anti-CD49b antibody (DX5) and EasySep mouse Biotin Positive Selection Kit II (Stemcell Technologies), followed by the isolation of CD200R3+cKit- cells by cell sorting. Library construction was conducted according to the TAS-seq protocol reported in the report by Shichino et al. bioRxiv 2022. Briefly, cells were indexed with Totalseq anti-mouse Hashtag antibody (BioLegend) for multiplexing. Reverse-transcription was conducted by using the BD Rhapsody according to the manufacturer's instruction. Preparation of cDNA library and hashtag library was performed as described previously (Chen CY et al. Cancer Immunol Res 2021). Sequencing librarie were prepared by utilizing NEBNext UltraII FS Library Prep Kit (New England BioLabs, USA).