Single cell transcriptome analysis of E15.5 and E17.5 mouse coronal sutures - all clusters

E15.5 and E17.5 coronal sutures were dissected from calvaria and individual cells isolated for scRNAseq. Transcriptome libraries for single cells were captured using 10X genomics Chromium Single Cell 3’ Library and Gel Bead Kit v2 following manufacturer’s guidelines. Sequencing for E15.5 coronal sutures was performed with Illumina’s HiSeq 3000/4000 PE Cluster Kit at the Children’s Hospital Los Angeles’ Molecular Genomics Core, and for E17.5 cells, PE sequencing was run on an Illumina HiSeq 4000 at the Oxford Genomics Centre Wellcome Centre for Human Genetics, Oxford achieving an average of ~150,000 mean reads per cell for E15.5 and ~92,000 mean reads per cell for E17.5.

本研究从颅盖骨（calvaria）中分离获取E15.5与E17.5时期的冠状缝（coronal suture），并解离得到单个细胞用于单细胞RNA测序（scRNAseq）。单细胞转录组文库的构建采用10X Genomics Chromium单细胞3'文库及凝胶磁珠试剂盒v2（Chromium Single Cell 3’ Library and Gel Bead Kit v2），严格遵循制造商的操作指南执行。其中，E15.5冠状缝样本的测序工作于洛杉矶儿童医院分子基因组学核心实验室完成，使用Illumina HiSeq 3000/4000双端聚类试剂盒（Illumina’s HiSeq 3000/4000 PE Cluster Kit）；E17.5细胞样本的双端测序则在牛津大学威康人类遗传学中心牛津基因组学中心的Illumina HiSeq 4000平台上开展，最终两组样本的平均每细胞读取数分别约为15万与9.2万。