Question: How does the performance of 10x Genomics’ RNase Inhibitor 40x compare to Protector RNase Inhibitor when used in 10x Genomics demonstrated protocols?
Answer: The RNase Inhibitor 40x (PN-1000887) sold by 10x Genomics has equivalent or greater activity than the Protector RNase Inhibitor (Sigma-Aldrich / MilliporeSigma, PN-3335402001 or PN-3335399001) and will provide comparable results when used with supported demonstrated protocols.
Below we highlight two side by side data demonstrations comparing results for both Chromium Single Cell Gene Expression and Visium Spatial Gene Expression applications.
Chromium demonstration
PBMCs from a healthy donor were acquired from AllCells and thawed according to 10x Genomics Cell Thawing Protocols for Single Cell Assays (CG000447). Cells were lysed following the Nuclei Isolation from Cell Suspensions & Tissues for Single Cell RNA Sequencing (CG000124) using either 10x RNase Inhibitor 40X (PN-1000887) or Sigma-Aldrich Protector RNase Inhibitor (PN-3335399001). Universal 3’ Gene Expression libraries were generated following the Chromium GEM-X Single Cell 3' v4 User Guide (CG000731). Comparable genes per cell (left) and UMAP clustering (right) were observed for both inhibitors.
Visium demonstration
Fresh frozen human lymph nodes were acquired from BioIVT. Tissue block preparation, sectioning, staining, and imaging followed the Visium HD Fresh Frozen Tissue Preparation Handbook (CG000763) using either 10x RNase Inhibitor 40X (PN-1000887) or Sigma-Aldrich Protector RNase Inhibitor (PN-3335399001). Probe hybridization, probe ligation, slide preparation, probe release, extension, and library construction followed the Visium HD Spatial Gene Expression Reagent Kits User Guide (CG000685). Comparable genes per 8 µm bin (left), and high spatial concordance between clusters (right) were observed for both inhibitors.
Products: Universal 3’ Gene Expression, Universal 5' Gene Expression, Epi Multiome ATAC + Gene Expression, Flex Gene Expression, Visium for Fresh Frozen, Visium CytAssist for FFPE, Visium CytAssist Spatial Gene and Protein, Visium HD