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Single Nuclei RNA-Seq application available on Nadia

Single nuclei RNA-Seq allows researchers to profile gene expression in difficult to isolate cells as well as archived tissue. To enable this application, Dolomite Bio has established high throughput sNuc-Seq on the Nadia Instrument. 

As part of this newly established high throughput sNuc-Seq protocol on the Nadia Instrument, Dolomite Bio will provide a tailored protocol for nuclei preparation based on previously published studies (Habib et al.  2017.). 

 

sNuc-Seq Diagram-Web

Following nuclei preparation, thousands of single nuclei from fixed or frozen tissue samples are encapsulated with single barcoded mRNA capture beads using droplet microfluidics on the Nadia Instrument. The Nuclei are lysed inside the droplets and their mRNA content is captured in the beads. 

Generating reproducible and cost-effective single nuclei data with Nadia

Using the Nadia Instrument, scientists can profile thousands of single nuclei consistently and reproducibly and also allow scientists to capture high numbers of genes with low doublet rates and low cost.

 

Genesvsreads_allminusextR1
Scatterplot_Dronc_Drop_markergenes_be

 

Efficient gene capture 

The number of genes and transcripts captured per nucleus ranged from 2000-25000 and 3500-4500, respectively. Both values are similar to previously published data. 

 

Correlation between sNuc-Seq and Drop-Seq 

A high correlation between average gene expression of single nuclei and cells can be observed. Marker genes specifically enriched in cells (red) and nuclei (blue) can clearly be distinguished. 

 

 

[1] Habib et al. “Massively parallel single-nucleus RNA-seq with DroNc-seq.” Nature methods 14.10 (2017): 955.