GSM7679612: spleen rep3; Rattus norvegicus; RNA-Seq
An untreated rat was euthanized, and the spleen was collected. Single-cell suspensions from the spleen were obtained by gently forcing the tissues through a 70 µm nylon mesh grid in RPMI-1640 (06261-65, Nacalai Tesque). After washing with the wash buffer, 15 mL of ACK buffer was added into the sample, and then incubated in a 37 °C water bath for 6 min to eliminate the splenic erythrocytes. The sample was washed with the wash buffer three times. Total RNA was prepared using ISOGEN II (311-07361, Nippon Gene Co., Tokyo, Japan) and purified using an RNeasy Plus Mini Kit (74136, Qiagen, Limburg, Netherland) with gDNA elimination using an RNase-Free DNase set (79254, Qiagen) according to the manufacturer's protocols. Total RNA was used to prepare RNA-Seq libraries with a NEBNext Ultra ll Directional RNA Library Prep Kit for Illumina.