Col2a1-Cre;Idh1LSL/+ embryonic growth plate - Bulk RNA-sequencing

As another method to determine differences between Idh1 mutant and control growth plates, we used Bulk RNA sequencing. Three E18.5 growth plates from our Idh1 mutant mice and three control mice were studied. Differential gene expression analysis revealed genes that were expressed in the distinct clusters, further supporting our single cell RNA data. In order to assess the Idh1 mutant expressing chondrocytes, we utilized Collagen2a1-Cre males crossed with Idh1 LSL/+ females. Growth plate chondrocytes from E18.5 distal part of femur and proximal part of tibia were collected and snap frozen. Total RNA was isolated and used for RNA-seq analysis. After sequencing and downstream analysis using Seurat, we observed distinct clusters of cells with Idh1 mutation Total RNA was isolated using Single Cell RNA Purification Kit and submitted to core facitly for library generation and sequencing