GSM8420851: AA 5+3 AML cells biol rep4; Mus musculus; RNA-Seq
Mice were euthanized at the point of maximum response of chemotherapy and femurs, tibias, pelvis and humeri were isolated and cleaned to remove muscle. Bones were then crushed using a pre-cooled mortar and pestle in ice-cold flow buffer (PBS containing 2% FBS), and the obtained BM cell suspension was passed through a 70 μm cell strainer. Red blood cells were removed by incubating cells for 5 minutes in ACK lysing buffer (Gibco) on ice, the remaining BM cells were washed and resuspended in ice-cold flow buffer containing the viability dye 7-aminoactinomycin D (7AAD; BD Biosciences). Viable cells (AML cells GFP+7AAD-) were sorted on a BD FACSAria II (BD Biosciences) with the sample collector cooled to 4°C. RNA was extracted with Trizole (Life Technologies, 15596018). RNA-seq libraries were generated using the Roche Kapa mRNA HyperPrep protocol according to the users' manual. Libraries were pooled and sequenced on the Illumina platform with 2x 38bp reads. RNA-seq libraries were generated using the Roche Kapa mRNA HyperPrep protocol according to the users' manual. Libraries were pooled and sequenced on the Illumina platform with 2x 38bp reads.