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10x chromium x9/11/2023 ![]() Please note : Researchers need to schedule with the UMGC the delivery of a dissociated suspension of live cells or nuclei: Discuss experimental goals, design, projected costs, and timeline.Ī planning meeting is needed to start a single-cell project to review your experimental goals and provide guidance on experimental design, as well as go over project logistics like turnaround estimates, scheduling, suspension buffer, and cell concentration requirements (varies based on assay). Low-depth QC sequencing of 10X libraries.įull-depth sequencing of 10X libraries. ![]() Capture, QC, and library prep.ġ0x Immune Profiling. The total amount of sequencing needed for a project is determined by the type of single-cell assay and the number of cells captured.ġ0x GEX scRNA-seq (3' or 5') standard capture.įeature Library Add-on (HTO, ADT, CMO, etc.), includes QC, and library prep.ġ0x Fixed GEX SinglePlex or multiplex, includes capture, QC, and library prep.ġ0x Multiome. Sequencing: The UMGC uses a MiSeq QC run to check the number of targeted single cells before going into full-scale sequencing on the NovaSeq 6000. The UMGC offers all the 10X single-cell assays to give researchers the widest variety of experiments. Up to eight samples can be processed per run allowing for the capture of up to 80,000 cells per run (cell capture efficiency ~65%). Two new commercially available kits improve the sample preparation workflow: 1) the Fixed RNA Profiling kit uses formaldehyde fixation to preserve samples at the point of collection, enabling sample batching for flexibility, and 2) the Nuclei Isolation kit enables analysis of frozen archival tissue.Ĭapture and Library Prep: We house the latest 10X Chromium X instrument. ![]() We can also arrange for a dry run to optimize your cell preparation for the workflow before going into full project with sequencing.Ĭell Prep: The 10X Genomics Cell Preparation Guide describes best practices and methods, but researchers should optimize their preparation protocol for their particular sample type. For cell samples needing sorting, the UMGC offers a MACSQuant Tyto cell sorter as a DIY instrument. See the above Guidelines tab for more info. WorkflowĬonsult: We request a project planning meeting to review experimental goals and provide guidance on experimental design. We also support library preparation and sequencing of cDNA samples derived from 10X’s Visium spatial solution. The UMGC offers a wide and growing list of 10X single-cell assays to support a broad range of applications, including gene expression, chromatin accessibility, cell surface proteins, immune clonotype, antigen specificity, and CRISPR screens. Additional support services are available on-campus through the University Flow Cytometry Resource (UFCR) and the Minnesota Supercomputing Institute (MSI).Support with experimental design and guidance on assay selection, including adding Feature Barcoding and Cellplex options, or our supported Cell Hashing and CITE-Seq protocols.10X libraries are sequenced on our production-scale NovaSeq 6000 for economical sequencing and project scalability.Latest 10X instrument for analyzing hundreds to hundreds of thousands of cells in a single run.Complete end-to-end workflow: cell suspension loading, 10X library creation, rigorous QC, sequencing, and analysis.The UMGC can handle a range of single-cell projects, from pilot experiments to the highest-throughput studies, and offers the full portfolio of validated 10X single-cell assays, to give researchers flexibility for any experimental strategy. The granularity of the proprietary Next GEM technology allows researchers to interrogate individual cells and unmask rare cell types that may not be seen in bulk RNA sequencing methods. The UMN Genomics Center expands single-cell research capabilities with the addition of 10X Genomics Chromium X, a next-generation microfluidic platform for single-cell analysis. ![]()
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