High-throughput advantage
The ICELL8 cx Single-Cell System (Cat. # 640188 or 640189) and plate-seq workflows both utilize well-based technologies. However the ICELL8 system's nanowell chips provide greater efficiency whether developing new chemistries or optimizing existing workflows.
By leveraging the capabilities of the ICELL8 cx system along with SMART-seq technology, the workflow provides a cost-effective solution preparing sequencing-ready libraries. Multiple plate-seq experiments can be combined into a single run using a nanowell chip.
Reduction in time and costs associated with preparing sequencing-ready libraries
| Comparison: Full-length scRNA-seq, 1,000-1,200 cells | |||||
| Number of experiments (plates/chips) | |||||
| ICELL8 cx | 1 | ||||
| Plate-seq | 13 | ||||
| Run-costs (pre-sequencing) | |||||
| ICELL8 cx | 1x | ||||
| Plate-seq | 16x | ||||
| Time to run and analyze | |||||
| ICELL8 cx | <3 days | ||||
| Plate-seq | ~2 weeks | ||||
Time estimation for 1,000-1,200 cells (SMART-seq on ICELL8 cx system)
| Day.Step # | Description | Time required | |
| Sample prep | 1 | Prepare cells |
2 hours |
| 1.1 | Harvest cells and stain | ||
| 1.2 | Wash and count | ||
| ICELL8cx SMART-seq application |
1.3 | Dilute cells and prepare source plate | |
| 1.4 | Dispense cells and image | ||
| 2 | Reverse transcription and cDNA library prep | 8 hours (Day 2) |
|
| 2.1 | Freeze chip at −80° C and thaw | ||
| 2.2 | Denature RNA | ||
| 2.3 | RT-PCR | ||
| 2.4 | Index1 dispense | ||
| 2.5 | Tagmentation dispense and incubation | ||
| 2.6 | Index2 dispense | ||
| Illumina Library Prep |
2.7 | Library PCR1 | |
| 3 | Nextera XT Library Prep | 2 hours (Day 3) |
|
| 3.1 | Bead purification (2 rounds) | ||
| 3.2 | Library PCR2 | ||
| 3.3 | Bead purification (1 round) | ||
| 3.4 | Final library QC |
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