Question: Can I use an alternative RNase inhibitor part number?
Answer: Different RNase inhibitors can have different enzyme mixes and enzyme activities so substituting the recommended part number is not recommended. Additionally, some RNase inhibitors have shown a reduction in cDNA yield or negative impacts on data quality. In the traces below, the recommended RNase inhibitor used in our demonstrated protocol, Protector RNase inhibitor* (Sigma, PN-3335399001), was replaced with an alternative, SUPERaseIn RNase Inhibitor (Thermo Fisher, PN: AM2694) and showed a significant reduction in cDNA yield when compared to Sigma Protector RNase inhibitor control.
Other RNase inhibitors tested and shown to negatively impact data quality:
- SUPERaseIn RNase Inhibitor
- Ribonucleoside Vanadyl Complex
It is strongly recommended to use the RNase Inhibitor part number listed in the Multiome ATAC + Gene Expression nuclei isolation demonstrated protocols (Sigma Protector RNase inhibitor, PN-3335399001).
Ambion™ RNase Inhibitor (AM2684 Thermo Fisher Scientific), RNaseOUT™ Recombinant Ribonuclease Inhibitor (10777019 Thermo Fisher Scientific) and, RiboLock RNase Inhibitor (Thermo Fisher, PN-EO0382) may be used as an alternative when Sigma Protector RNase inhibitor is not available. Note: there was limited testing of these inhibitors for Multiome ATAC + Gene Expression, so use with caution.
*If using the Chromium Nuclei Isolation Kit, the use of alternative RNase inhibitors is unsupported. Utilize the provided and validated RNase Inhibitor for best results.
Products: Single Cell Multiome ATAC + GEX