GSM5608830: L. major promastigote parasite infected 05; Mus musculus; RNA-Seq
At 4 week, with or without infection, mice ears were excised and enzymatically digested using 0.25 mg/ml Liberase and 10 mg/ml Dnase-I in incomplete RPMI 1640 for 90 minutes at 370 C. After digesting, ears were minced manually to obtain cellular content in single cell suspensions. The total RNA was isolated from the cell lysate of naive and infected ears by using RNeasy plus mini kit according to the manufacturer's instruction. cDNA libraries were prepared from total RNA using Lexogen's QuantSeq 3' (FWD) mRNA-Seq Library Prep Kit for Illumina following the manufacturer's protocol. Library fragment sizes were validated using a Fragment Analyzer and quantified by a Qubit fluorometer. Equal amounts of each library were pooled for sequencing on the NextSeq 500 platform using a high output flow cell to generate approximately 25 million 75bp SE reads per sample.