Chapman Alec R, He Zi, Lu Sijia, Yong Jun, Tan Longzhi, Tang Fuchou, Xie X Sunney
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, 02138, United States of America; Graduate program in Biophysics, Harvard University, Cambridge, MA, 02138, United States of America.
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, 02138, United States of America.
PLoS One. 2015 Mar 30;10(3):e0120889. doi: 10.1371/journal.pone.0120889. eCollection 2015.
Recently, Multiple Annealing and Looping-Based Amplification Cycles (MALBAC) has been developed for whole genome amplification of an individual cell, relying on quasilinear instead of exponential amplification to achieve high coverage. Here we adapt MALBAC for single-cell transcriptome amplification, which gives consistently high detection efficiency, accuracy and reproducibility. With this newly developed technique, we successfully amplified and sequenced single cells from 3 germ layers from mouse embryos in the early gastrulation stage, and examined the epithelial-mesenchymal transition (EMT) program among cells in the mesoderm layer on a single-cell level.
最近，基于多次退火和成环的扩增循环（MALBAC）技术已被开发用于单个细胞的全基因组扩增，该技术依靠准线性而非指数扩增来实现高覆盖率。在此，我们将MALBAC技术应用于单细胞转录组扩增，该技术能持续提供高检测效率、准确性和可重复性。利用这项新开发的技术，我们成功地对处于原肠胚形成早期阶段的小鼠胚胎三个胚层的单细胞进行了扩增和测序，并在单细胞水平上研究了中胚层细胞间的上皮-间质转化（EMT）程序。
