Nuc-Seq with EdU-Mediated Labeling of Proliferating Cells
Div-Seq is a single-nucleus RNA sequencing technique that improves upon Nuc-Seq by incorporating 5-ethynyl-2ê-deoxyuridine (EdU) labeling to identify dividing cells during their different cell stages (Habib et al., 2016). EdU labeling also enables identification of different cell types in complex tissue samples and rare cell populationsduring FACS.
Briefly, samples are labeled in vivo with EdU, dissected, and fixed before isolation into single nuclei. Individual nuclei are tagged fluorescently and sorted by FACS. From this step, the procedure follows the Nuc-Seq method: cDNA synthesis is performed using the Smart-seq2 protocol, while the cDNA library is prepared with Tn5 transposase_mediated tagmentation.
Similar methods: Nuc-seq, snRNA-seq
- Tracks transcriptome dynamics in single nuclei
- Detects rare cell populations
- Compatible with fresh, frozen, or fixed sample types
- EdU labeling gives unbiased identification of different types of dividing cells and their current stage in the cell cycle
- Mild nuclear dissociation technique minimizes gene expression changes commonly seen in protease-mediated dissociation
- Excludes any information from cytoplasmic RNA
Illumina Library prep and Array Kit Selector
Wagner A., Regev A. and Yosef N. Revealing the vectors of cellular identity with single-cell genomics. Nat Biotechnol. 2016;34:1145-1160
Goncalves J. T., Schafer S. T. and Gage F. H. Adult Neurogenesis in the Hippocampus: From Stem Cells to Behavior. Cell. 2016;167:897-914
Habib N., Li Y., Heidenreich M., et al. Div-Seq: Single-nucleus RNA-Seq reveals dynamics of rare adult newborn neurons. Science. 2016;353:925-928