GSM7527555: Ppia heterozygous replicate 2; Mus musculus; RNA-Seq
Wild-type HSPCs were isolated from the hind limb long bones of 10-12 month-old female mice. c-Kit+ cells were stained and magnetically isolated from the lineage-depleted cell suspension using the EasySep mouse CD117 (c-Kit) positive selection kit (Stem Cell Technologies), following the manufacturer's instructions. RNA extraction was carried out with the RNeasy Plus Mini kit with genomic DNA Eliminator columns (Qiagen) in combination with on-column DNaseI digestion (Qiagen), according to the vendor's protocol. Total RNA was quality-checked using Agilent RNA 6000 Pico kit (#5067-1513) and the Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, USA). RNA with DV200 greater than 66% was used for library preparation. Total RNA-seq libraries were generated and prepared for multiplexing on the Illumina platform with the SMARTer Stranded Total RNA-Seq Kit v2 (#634418, Takara Bio, Japan) and the Unique Dual Index Kit (#634752, Takara Bio, Japan) following the manufacturer's instructions. The libraries were sequenced on an Illumina NovaSeq 6000 System (Illumina, Inc., USA) with 150 nt paired-end sequencing.