GSM6807992: bone, 2000µɛ loaded, 0 h post-culture, donor5; Homo sapiens; RNA-Seq
Bone explants were pulverized using a 6775 Freezer/Mill cryogenic grinder (SPEX SamplePrep, Metuchen, NJ, USA). Explants were grinded (rate: 10 cycles/sec; 20 impacts/sec) for 2 min in liquid nitrogen. After pulverization, the TRIzol/bone powder mixture was incubated for 1 h in a shaking waterbath at 37°C. Isolation and purification of RNA was performed using RNeasy Midi columns (Qiagen, Hilden, Germany) or Zymo-Spin IIICG columns (Direct-zol RNA MiniPrep Plus, Zymo Research, Irvine, CA, USA). cDNA libraries for RNA-seq were prepared using KAPA mRNA Hyperprep kit (Roche Sequencing Solutions, Indianapolis, IN, USA).