title: GSM8525415: AB6638_SB181; Homo sapiens; RNA-Seq
library_construction_protocol: Synovial fluid mononuclear cells and blood mononuclear cells were isolated within maximum 4 hours of sample collection. Synovial fluid samples were first filtered by 100-micron strainer into 50 ml tube and diluted 1:1 with ice cold FACS buffer (2mM EDTA, Ph 8.0, 0.5% bovine serum albumin in PBS). Blood samples were diluted 1:1 with ice cold FACS buffer. Synovial fluid and blood mononuclear cell separation was performed using density centrifugation media (Ficol-paque, GE Life Sciences) in a 1:1 ratio with the diluted synovial fluid or blood sample. Centrifugation (460g, 25min) was performed at 10◦C and the mononuclear cells were carefully aspirated and washed with ice cold FACS buffer. After a short centrifuge (360g, 5min), we resuspended the cell pellet in cryopreservation media (FCS+10%DMSO) and stored the samples in cryotube at -80◦C. Prior to single-cell sorting, samples were quickly defrosted at 37◦C bath and washed with heated (37 ◦C) RPMI-1640. We resuspended the cells in FACS buffer for staining and sorting. 3' end mRNA libraries were prepared for sequencing using SPID-seq (Yofe et al, Cancer Discovery 2023)