GSM9405592: KA13_SF_time_OP0_Control; Equus caballus; RNA-Seq
The fluid was digested with 0.3 mg/mL hyaluronidase (Sigma, St. Louis, MO, USA) by incubation in a bead bath for 15 minutes at 37 ºC. Next the sample was centrifuged at 750 x g for 10 minutes at 8 ºC. The remaining cell pellet was washed with PBS and centrifuged at 500 x g for 10 minutes at 8 ºC. The cell pellet was resuspended in about 1 to 2 mL of PBS and transferred into a 2 mL microcentrifuge tube. The sample was then centrifuged for 500 x g for 15 minutes. The supernatant was discarded, and the cells were lysed in RLT buffer (Qiagen, Germantown, MD, USA) with 1% ß-Mercaptoethanol (BME; EMD Millipore Corp, Billerica, MA, USA) and stored at -80 ºC until RNA extraction. Library preparation was conducted on synovial fluid samples using the SMARTer® Stranded Total RNA-Seq Kit -Pico input (Takara Bio USA, Inc.)