Single nuclei RNA-Seq (sNuc-Seq) is a novel methodology which uses isolated nuclei instead of whole cells to profile gene expression in cells which are difficult to isolate, as well as archived tissue. Using droplet microfluidic technology this enables users to profile thousands of nuclei at low cost and at high throughput.
Nuclei can usually be rapidly and easily isolated from lightly fixed, frozen tissues and archived without the extended incubation and processing required for isolating single cells. The rapid isolation of nuclei from complex tissue will allow researchers to acquire transcriptomes that are relatively unperturbed by the isolation procedure
A recently published method, ‘DroNc-Seq’ (1) by Habib et al. described the use of single nuclei as a proxy for whole cells to generate thousands of single nuclear transcriptomes. This allows the use of cells that cannot be readily dissociated or are fixed. This method which, has been adapted for use on the Nadia platform, is called single nuclei RNA sequencing (sNuc-Seq).
Like the Drop-seq method, sNuc-seq uses droplet microfluidics to encapsulate single nuclei with uniquely barcoded mRNA capture beads. Following downstream processing and sequencing, the transcriptomes of thousands of single nuclei can be analysed, and unique cell types identified.
Previously research was limited in cells that were difficult to isolate or archived tissue, preventing the discovery of new cell types or vital information about disease and immunity. With the introduction of sNuc-Seq technologies, new insights and discoveries are now be possible into various research areas, including:
Dolomite Bio have established high throughput sNuc-seq on the fully automated Nadia instrument. This now allows researchers to easily generate high quality single nuclei sequencing libraries as originally described by Habib et al. 2017 (1). Habib et al. “DroNc-Seq: Deciphering cell types in human archived brain tissues by massively-parallel single nucleus RNA-Seq.” bioRxiv (2017): 115196.