The single cell multiome approach provides for simultaneously profiling of open chromatin regions and gene expression in the same single nuclei. The Next GEM Single Cell Multiome ATAC + Gene Expression kit from 10x Genomics captures nuclei which have been incubated with a transposase to preferentially fragment open regions of chromatin. Nuclei are partitioned into nanoliter-scale Gel Beads-in-emulsion (GEMs) to capture mRNA (poly(dT) primers) and DNA fragments (spacer sequence) and incorporate a 10x barcode allowing for transcripts and DNA fragments to be associated with specific cells.
Key features of the service:
- Joint consultation available with Genomics Core and Bioinformatics Core.
- Cell counting with K2 Cellometer and cell suspension loading.
- Chromium X instrunment to process cells with latest GEM-X reagents.
- 10x library construction and rigorous library QC.
- Sequence generation performed on a NovaSeq X Plus platform providing economical sequencing and project scalability.
- Biosafety level 2 containment available for cell collection.
Additional documentation and resources are available on the 10x Genomics support site
A meeting is required with core staff to initiate a single cell project. The meeting will discuss experimental goals as well as project turnaround times, collection scheduling, cell concentration requirements, and cell/nuclei quality requirements. Arrangements for a meeting to discuss a project can be made by sending an email to Nathan Bivens, Genomics Core director.
All projects must be initiated with a quote, and quote accepted, before a collection can be scheduled. .
The 10x Genomics Single Cell Calculator may be used to estimate the amount of sequence required for a project.
General Considerations:
- Submit cells in Eppendorf DNA Lo Bind Tubes, 2.0 ml (Cat. No. 022431048)
- Must minimize presence of cellular aggregates, dead cells, non-cellular nucleic acids and reverse transcription inhibitors.
- On the scheduled day of submission, core staff should be notified 1 hour prior to cells/nuclei being delived. It is best practice to provide periodic updates to core staff on the day of collection.
Submission Requirements:
| Nuclei Concentration | Volume | Required Buffer | Special Instructions |
| 3,200 - 8,000 nuclei/ul | 30 - 100 ul | Diluted Nuclei Buffer *provided in kit |
Review the 10x article on how to assess nuclei quality. |
| Sequence Depth/Coverage* | Read Type | Index Type | Read Length | ||
| 25,000 read pairs/nucleus (ATAC) | Paired end | unique dual-index | 50 bases | ||
| 20,000 reads/nucleus (GEX) | Paired end | unique dual-index | 100 bases | ||
* minimum read count recommended by MUGTC; increased sequencing depth may be desired for some studies