The RNA-Seq system includes all the components required to encapsulate a cell sample. It uses the same reagents and beads described in the Drop-seq protocol, simplifying adoption and increasing reliability.
- Generates more than 6,000 single cell libraries in a 15 minute run
- Quick and reproducible encapsulation of single cells
- Doublet rate less than 0.5 % at limiting dilution of 1:20 cells per droplet.
- Second channel allows co-encapsulation with oligonucleotide-barcoded beads and lysis buffer
- Automation and integrated droplet analysis via PC software
- Premium service – installation and training included
The Single Cell RNA Seq System is particularly suitable for the Drop-seq protocol as described in Macosko et al., 2015, Cell 161, 1202–1214. The system includes Single Cell RNA Seq chips, plus the connectors and tubing required to use the chips with Dolomite Bio Pressure pumps and flow sensors. A High Speed Camera and Microscope is included to allow droplet production to be visualised.
The Dolomite Bio system offers unique capability which is not available elsewhere. The unique features include:
- Single-channel pump with integrated reservoir, which allows compact setup, and also allows pumps to be used separately for different applications
- 10 bar pressure pump capability, allowing high flow rates
- Safe pressure vessel design, using self-locking reservoir lids and pressure vessel rated glassware, to eliminate the risk of vial fracture in operation
- Scalable to high-throughput, using modular reservoirs and high-throughput chips
- Vacuum and pressure capability in the same pump for maximum flexibility
- Modular flow sensor design, allowing cost-effective use of a range of flow sensors
- Common connector format, allowing modular assembly of multiple chips from a small number of parts
- High chemical, thermal, and pressure resistance, with reusable microfluidic chips
The Dolomite Bio Single Cell RNA Seq chip offers unique benefits over standard PDMS chips, including higher reagent throughput, long life, and ease-of-use.
To encapsulate a sample, the user loads a cell suspension into an externally agitated microcentrifuge tube and loads a bead suspension into a sample loop via a syringe.
The droplets are collected in an output reservoir at approximately 2,800 droplets per second. Complete processing of a sample occurs 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 rate and pressure.
The hardware is controlled by the Flow Control Centre software.
An RNA Seq system includes:
Hardware (3 pressure pumps, 3 flow sensors, sample loop and injection valve, microscope and camera), control software (Flow Control Centre Advanced), accessories & consumables (chips, connectors, valves and pipes), reagents (emulsion oil) 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