GSM7679608: whole blood cells rep4; Rattus norvegicus; RNA-Seq
An untreated was placed under anesthesia, and blood was collected from the heart by cardiopuncture. The blood was washed once with wash buffer once. After removing the supernatant, 15 mL of ACK buffer was added to the sample and then incubated in a 37 °C water bath for 6 min to eliminate erythrocytes. After incubation, 25 mL of the wash buffer was added to the sample, which was then centrifuged at 400 g at 4 °C for 5 min. The incubation and wash operations were repeated twice. After the last wash, 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.