Single Cell Assay & Sequencing
The Single Cell Sequencing facility is located on the 2nd floor of the Barshop Building (Rm 2041). The core provides cost-effective and time-efficient access to cutting-edge genomic technologies and expert assistance with experimental design and data analysis. We offer two different platforms with the ability to sequence RNA, DNA and protein on a single cell level. The services include 3’ gene expression (V3.1, V3.1 HT, V4 GEM-X, V4 GEM-X Multiplex), 3’ gene expression with cell surface protein expression, 3’ gene expression multiplexing with cell hashing or multiome, 5’ gene expression, 5’ gene expression with immune profiling, 5’ gene expression with cell surface protein expression, Flex Gene Expression, Single Cell transposase-accessible chromatin using sequencing (ATAC-seq), Single cell multiome with ATAC & 3’ gene expression.
We also provide service for Visium spatial gene expression library prep through collaboration with Shock center Pathology Core, including Visium spatial fresh frozen or FFPE tissue gene expression library prep.
Appointments are required for the 10X single cell service, and we kindly request advance notice (at least 1 week) due to the specialized preparations involved. Please follow the 10x Genomics Procedures for all the single cell service.
10x Genomics Procedures:
- Pre-consultation: This is mandatory for all the single cell assays. These are the topics we would discuss during consultation: sample type and number, single cell assay type, single cell or nuclei preparation, sample quality, anticipated cells numbers, sequence saturation concept, workflow of the whole project, price quote, and collaborations on the data analysis.
- Email: confirm the submission form and reserve appointments for the assay day.
- Assay Day: bring the sample on ice, and printed submission form. We will check the quality of the samples. Any low-quality samples will not be processed. For other assays, protocols may be modified accordingly.
- Transfer sequencing results and discuss data analysis part.
Equipment: 10X Genomics Chromium X
The latest release from 10X Genomics for single cell encapsulation, the Chromium X allows for capture of up to 20,000 cells per lane, with even higher numbers for fixed cells. Methods are available for 3’ and 5’ library prep as well as 3’ fixed cells from human and mouse. This new platform combines the well-established 10X methodology with new higher throughput methods to increase cell target number and decreasing doublet rates.
