GSM5769232
Primary epiphyseal chondrocytes were harvested from five-day-old male wildtype (n=3) and Girk3-/- (n=3) mice. Chondrocytes were digested for one hour in 0.5mg/mL collagenase II in DMEM culture medium, then digested overnight in 3mg/mL collagenase II in DMEM culture media. Cells were isolated the next day, washed in ice-cold PBS three times, then resuspended in Trizol for RNA extraction. Total RNA was extracted using phenol-chloroform. TruSeq RNA method was used for analysis of polyadenylated mRNAs that were selected using oligo dT magnetic beads. TruSeq Kits were used for indexing to permit multiplex sample loading on the flow cells of an Illumina HiSeq 2000 sequencer. Gene expression was normalized to one million reads and corrected for gene length (reads per kilobasepair per million mapped reads, RPKM).