title: GSM8717347: multiplexed CSF cells2; Homo sapiens; RNA-Seq
library_construction_protocol: CSF was collected on ice and processed according to a written standard operating procedure by investigators blinded to diagnoses, clinical, and imaging outcomes. Aliquots were assigned alphanumeric identifiers and centrifuged for 10 minutes at 300g at 4°C within 30 minutes of collection. The pelleted CSF cells were processed by flow cytometry or processed for single cell RNA sequencing. Fresh CSF cells were collected as described in sample processing. Cells were labelled with 79 Totalseq B antibodies (Biolegend, USA) and washed with PBS with 1% Bovine Serum Albumin (Thermo Fisher Scientific, USA). Labeled cells were fixed by using fixed RNA feature barcode kit (10x Genomics, USA) and stored at -80 degree.  Cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq) method was used to analyze transcriptome and surface protein information based on  single-cell readout. After probe hybridization, gene expression library and surface protein library were constructed by using Chromium Next GEM Single cell 3` LT Reagent kits v3.1 (Dual index) (10X Genomics, USA) following the protocol (CG000400). Single-cell fixed RNA libraries and single-cell feature barcode libraries were sequenced  on a NextSeq 2000 P3 28 cycle + 90 cycle asymmetrical run.  Demultiplexing yielded over 27 million reads per sample when allowing for 1 mismatch per index adapter.