Question: Where are the stopping points in the Gene and Protein Expression workflow?
Answer: Below are the stopping points throughout sample preparation and library construction:
- After tissue baking: Tissue baked at 60°C for 2 hours can be stored in a room temperature desiccator for up to 2 weeks
- Before or after H&E imaging: Coverslipped tissue is stable for 24 hours at 4°C
- During the assay:
- The User Guide workflow occurs over three days (post-tissue baking). The first day includes staining and Hybridization. The second day includes antibody labeling. The third day includes completing CytAssist-mediated probe and antibody tag transfer, on-slide extension, and off-slide library construction steps.
- Stopping points have been built into the third day of the workflow to maximize flexibility.
- After completing the last Post-Ligation Wash, samples can be stored in 2X SSC Buffer at at 4°C for ≤24h
- After completing the last Post-Antibody Incubation Wash, samples can be stored in 1X PBS at 4°C for ≤2h*
- *indicates that pause point is unvalidated but low risk
- After completing Extension, samples can be held at 4°C for ≤2h in the thermal cycler
- After completing the Pre-Amplification SPRIselect Cleanup, samples can be stored at 4°C for ≤72h or at -20°C for ≤4 weeks
- After completing the Sample Index PCR and SPRIselect Cleanup (for both Gene Expression and Protein Expression), samples can be stored at -20°C for long-term storage
Products: Visium CytAssist Gene and Protein