Journal: Frontiers in Neuroscience
Article Title: Exploring the Complexity of Cortical Development Using Single-Cell Transcriptomics
Figure Lengend Snippet: Current workflow of single-cell technologies to study cortical development. Step1. Biological systems to study brain development. Upper panel shows in vivo mouse embryonic brain and below panel indicates in vitro human brain organoid which is commonly used for the single-cell neurogenesis studies. Step2. Cell isolation methods. Individual cells can be isolated using FACS, Microfluidic ChIP, or Drop-seq approaches. Step3. Library preparation. The common protocols include polyA+ mRNA capture, reverse transcription, cDNA amplification using PCR, and tagmentation. Step4. Sequencing of the library. Step5. Computational analysis. After the preprocessing of sequencing reads, visualization using t-SNE, unsupervised clustering, and correlation analysis with bulk RNA-seq is followed to identify subtypes of cells and characterize their identities.
Article Snippet: In case of Smart-seq, commercially available Smart-seq kit (Clontech) is used to generate full-length double-stranded cDNA which is converted into sequencing libraries by tagmentation (Nextera, Illumina) (Ziegenhain et al., ).
Techniques: In Vivo, In Vitro, Cell Isolation, Isolation, FACS, Chromatin Immunoprecipitation, Amplification, Polymerase Chain Reaction, Sequencing, RNA Sequencing Assay