Chromium 3’

Owner: Gaby

Overview

Our lab houses a Chromium platform from 10x Genomics for single-cell and single-nucleus RNA sequecing experiments. *For lab members who will be involved in performing or assissting collaborators with these experiments, please review the scRNAseq SOP document, which covers how to set up a new experiment, sample preparation, data generation and handling, and instrument handling.

*This SOP applies specifically to Universal 3’ kits from 10x Genomics

Materials

Kits for Universal 3’ single-cell or single-nucleus RNA sequencing experiemnts can be purchased directly from 10x Genomics. All other necessary reagents are specified within the specific 10x Genomics protocols.

Protocol

Protocols for single-cell experiments can be found on the 10x Genomics website. For our experiments, we typically utilize the Chromium GEM-X Single Cell 3’ v4 Gene Expression User Guide or the user guide for GEM-X Universal 3’ Gene Expression v4 4-plex Reagent Kits for performing On-chip Multiplexing (OCM).

Results / Output

In the wet-lab, a single-cell experiment ends with the construction of a library. Libraries are quantified by qPCR, pooled, and submitted to Novogene for sequencing. Details on sequencing setup and submission can be found within the Sequencing SOP.