Landskron Lisa, Bonnay Francois, Burkard Thomas R, Knoblich Jürgen A
IMBA - Institute of Molecular Biotechnology of the Austrian Academy of Science, Vienna Biocenter (VBC), Vienna, Austria.
Bio Protoc. 2020 Nov 5;10(21):e3809. doi: 10.21769/BioProtoc.3809.
Cell-type specific transcriptional programs underlie the development and maintenance of organs. Not only distinct cell types within a tissue, even cells with supposedly identical cell fates show a high degree of transcriptional heterogeneity. Inevitable, low cell numbers are a major hurdle to study transcriptomes of pure cell populations. Here we describe DigiTAG, a high-throughput method that combines transposase fragmentation and molecular barcoding to retrieve high quality transcriptome data of rare cell types in . The protocol showcases how DigiTAG can be used to analyse the transcriptome of rare neural stem cells (type II neuroblasts) of larval brains, but can also be utilized for other cell types or model systems.
细胞类型特异性转录程序是器官发育和维持的基础。不仅组织内不同的细胞类型,甚至具有假定相同细胞命运的细胞也表现出高度的转录异质性。不可避免的是,低细胞数量是研究纯细胞群体转录组的主要障碍。在这里,我们描述了DigiTAG,一种高通量方法,它结合转座酶片段化和分子条形码来检索稀有细胞类型的高质量转录组数据。该方案展示了DigiTAG如何用于分析幼虫大脑中稀有神经干细胞(II型神经母细胞)的转录组,也可用于其他细胞类型或模型系统。