GSM4909030: Pdgfra+/+_2; Mus musculus; RNA-Seq
Secondary palatal shelves (PS) and maxillary processes (MxP) were dissected on ice and the overlying ectoderm removed by digestion with 20 mg/mL trypsin for 15-20 min at 4°C. PS and MxP were subsequently rinsed in 10% fetal bovine serum for 1 min followed by 1x phosphate buffered saline (PBS). Excess PBS was removed and PS and MxP were submerged in 200 uL Invitrogen RNAlater stabilization solution for overnight storage at 4°C. The following day excess RNAlater was removed and the samples were stored at -80°C. Total RNA was simultaneously isolated from all samples using the Qiagen RNeasy Mini Kit according to the manufacturer's instructions. PolyA selection was performed and sequencing libraries were synthesized using NuGen Universal Plus mRNA-Seq with NuQuant kit at the University of Colorado Genomics and Microarray Core. Libraries were sequenced on an Illumina NovaSeq6000.