GSM5387044: RNA-seq_osteoblasts_Runx2_cHom_KO_rep2; Mus musculus; RNA-Seq
Rib chondrocytes were manually dissected from the neonatal mouse ribs. The ribs were initially digested in 2 mg/mL Pronase (53702, EMD Millipore Corporation) for 30 min at 37 °C, washed three times with phosphate buffered saline (PBS), and then digested in 0.2% collagenase D (1088858, Roche Applied Science) for 3 h at 37 °C. Calvarial osteoblasts were manually dissected from osteoblast-enriched partial calvarias consisting of the frontal bone and a rostral part of the parietal bone. The partial calvarias were digested in five changes of 0.1% collagenase D and 0.2% dispase for 10 min each at 37 °C. For RNA-seq with rib chondrocytes sorted from Col2a1-ECFP and Col10a1-mCherry mice at P1, 100 ng of total RNA was subjected to analysis. RNA libraries were constructed using a Tru-Seq Illumina kit (RS-122-2001, Illumina) with some modifications. For RNA-seq for with Runx2_cHet_KO cells and Runx2_cHom_KO cells from calvaria, 1.7 ng of total RNA was subjected to the analysis. RNA libraries were constructed using a SMARTer Stranded Total RNA-Seq Kit v2 Pico Input Mammalian (634411, Takara).