The RNA-Seq system is ideal for performing the Drop-seq protocol, simplifying adoption and increasing reliability. Cell samples can rapidly be co-encapsulated with lysis buffer and oligonucleotide-barcoded beads to generate a great number of single cell libraries.
- High throughput: More than 6,000 single cell libraries in a 15-minute run.
- Flexible open system: Full control of pressures, flow rate and buffers/reagents.
- High quality results: Doublet rate less than 0.5 % and extremely monodisperse emulsion (CV <5%).
- Easy to use: Off the shelf advanced microfluidic products.
- Minimal damage to beads: The Sample Loop and Injection Valve module uses no harsh mechanical stirring avoiding bead damage.
- Installation & training: The exceptional service includes onsite installation and training by our experienced Applications team.
- Integrated droplet analysis via PC software: Real time images of the dropletization enable quality control of droplet formation and development of new protocols.
- Optional modified RNA-Seq chip to perform DroNc-Seq: Enables users to profile thousands of nuclei at low cost and at high throughput
The Single Cell RNA-Seq System is particularly suitable for the Drop-seq protocol and reagents as described in Macosko et al., 2015, Cell 161, 1202–1214. Furthermore, the system is flexible and open making it accessible for the use of a wider range of reagents and the development of own protocols.
A cell suspension is loaded into an agitated remote reservoir and beads are injected into a sample loop ensuring an even loading of both cells and beads. Droplets are collected in an output reservoir at approximately 2,800 droplets per second allowing for the capture of over 6000 single cells in 15 minutes.
Highly monodisperse droplets are produced with precise control using the pressure pumps. The flow sensors provide real time display of micro-scale flow rates and allow for accurate adjustment of flow using the Flow Control Centre software. Droplet production can be monitored with the High Speed Camera and Microscope System, allowing for real-time control over quality of droplet formation.
The Dolomite Bio system offers unique capability which is not available elsewhere, including:
- Remote reservoir for stirring of cells to keep them in suspension without using neutral buoyancy medium
- Sample loop and injection valve ensures equal loading of beads into droplets and uses no harsh mechanical stirring thereby preventing broken or damaged beads
- Scalable to high-throughput, using modular reservoirs and high-throughput chips
- High quality Single Cell RNA-Seq chip which offers unique benefits including higher reagent throughput, long life, and ease-of-use
A Single Cell RNA-Seq system includes:
Hardware (3 pressure pumps, 3 flow sensors, sample loop and injection valve, compressor, monitor, microscope and camera), control software (Flow Control Centre Advanced), accessories & consumables (chips, connectors, valves and pipes) and installation & training.
- Material encapsulated: Cells and oligo barcoded beads
- Monodispersity of droplets: Extremely monodisperse (CV <5%)
- Droplet diameter: Typically 88µm
- Droplet volume: Typically 357 pl
- Sample volumes: 500 µl cell suspension + 500 µl reagent mix/bead suspension
- Pump pressure range: 0-10 bar
- Flow rate range: Dependent upon pressure and flow restriction but achievable rates from nl/min to ml/min
- Droplet frequency: Typically 2.8 kHz
- Quantitative analysis of droplet size is achievable at low flow rates – typically below 1 kHz – and qualitative evaluation at production speeds of 2-15 kHz or higher, depending on droplet size
- Wetted materials: Glass, FEP, FKM