Chapter title |
A Modified SMART-Seq Method for Single-Cell Transcriptomic Analysis of Embryoid Body Differentiation.
|
---|---|
Chapter number | 435 |
Book title |
Embryonic Stem Cell Protocols
|
Published in |
Methods in molecular biology, October 2021
|
DOI | 10.1007/7651_2021_435 |
Pubmed ID | |
Book ISBNs |
978-1-07-162436-4, 978-1-07-162437-1
|
Authors |
Zheng, Jianqun, Ye, Ying, Xu, Qiushi, Xu, Wei, Zhang, Wensheng, Chen, Xi |
Abstract |
Embryoid bodies (EBs) are aggregate of cells that contain three embryonic germ layers. They can be formed by direct differentiation from pluripotent embryonic stem cells (ESCs), which serves as a useful model for understanding early embryo development. Due to the mixture of different cell types, it is necessary to investigate EBs at the single-cell level. Here, we describe a robust and straightforward method for single-cell gene expression profiling during mouse EB differentiation from mouse ESCs (mESCs). The protocol is modified from a widely used method in the SMART-seq family, which only requires standard molecular biology techniques and lab equipment. It allows for accurate 3' counting of transcript at the single-cell level, which helps reveal cellular identities during EB formation. Combined with perturbation experiments, the method provides an opportunity for mechanistic studies of embryo development at the single-cell level. |
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