Question: Which version of the Visium Spatial Gene Expression User Guide is supported for use with the Targeted Gene Expression assay?
Answer: When preparing Visium Spatial Gene Expression libraries for use with the Targeted Gene Expression assay, it is essential to use the most recent version of the Visium User Guide (CG000239 Rev F). Visium Spatial Gene Expression libraries prepared using earlier versions (Rev A - Rev C) are not compatible with the Targeted Gene Expression assay. See: Why were the Sample Index PCR conditions updated for Visium Spatial Gene Expression library preparation?
If you have already generated Visium Spatial Gene Expression libraries using an earlier User Guide version (Rev A - Rev C), we recommend re-preparing the libraries. If stored cDNA is available for these samples, this cDNA can be used to re-prepare libraries using the updated library preparation protocol in User Guide Rev F. Use stored cDNA (from the “cDNA cleanup” step 3.3n of CG000239 Rev C or step 4.3n of CG000239 Rev A-B) as input into the “Visium Spatial Gene Expression Library Construction” step (Step 4 of CG000239 Rev F).
The Library Construction Kit (PN-1000190) can be purchased as a standalone kit to re-prepare the Visium Spatial Gene Expression libraries from left-over cDNA. We have successfully re-prepared libraries from cDNA stored for up to one year at - 20 °C. These re-prepared Visium Spatial Gene Expression libraries can be used as input into the Targeted Gene Expression assay.
The Targeted Gene Expression assay is not compatible with libraries prepared using the Visium for FFPE assay.
Product: Visium for fresh-frozen, Targeted Gene Expression