|Public on Nov 27, 2018
cell type: Immortalized bone marrow derived macrophages
genotype: wild type
|Cells were infected by lenti-virus containing GRA15II gene and selected by applying puromycin in culture medium
|Immortalized bone marrow derived macrophages were cultured in DMEM containing 10% FBS, 1% glutamine, 1% streptomycin-penicillin on 10 cm dish for further experiments.
|Supernatant was removed and cells were covered with 350ul Buffer RLT at the end of treatment. Total RNA was extracted following the strandard protocol of RNeasy Mini kit (Qiagen).
RNA libraries were prepared for sequencing using standard Illumina protocols
|Illumina NovaSeq 6000
|RNA sequencing libraries were generated using NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB), and sequenced on an Illumina Novaseq platform.
Illumina Casava 1.8 software used for basecalling and raw reads were gained.
After eliminating low-quality reads and reads with adaptors, clean reads were mapped to mouse genome reference sequence (GRCm38/mm10 Ensembl release 81) using HISAT2.
Gene expression was quantified as reads per kilobase of coding sequence per million reads (RPKM) algorithm.
Supplementary_files_format_and_content: tab-delimited text files include ensemble gene ID and RPKM values for each Sample
|Nov 26, 2018
|Last update date
|Nov 27, 2018
|NO.38 ROAD XUEYUAN, DISTRICT HAIDIAN, BEIJING, CHINA, 北京市海淀区学院路38号北京大学医学部
|mRNA-seq of immortailized bone marrow derived macrophages (iBMDM) overexpressing GRA15II