Question: Can I use an alternative to the KAPA SYBR Fast qPCR Master Mix (KK4600) for Cycle Number Determination during the Visium Gene Expression workflows?
Answer: We DO NOT recommend using an alternative qPCR Master Mix for Cycle Number Determination. Only the KAPA reagent has been validated with the Visium and Visium for FFPE Gene Expression assays. The KAPA SYBR Mix contains a fast DNA polymerase, allowing for Cycle Number Determination within 25 cycles.
In the United States, the KAPA SYBR Fast qPCR Kit can be ordered directly from Roche (Universal kit PN: KK4600, or kits with alternative volumes KK4601- 5ml, and KK4602- 10ml). Alternatively, the KAPA SYBR FAST qPCR Master Mix (KM4101) in the KAPA Library Quant Kit (Universal kit PN: KK4824) can also be used for the cycle number determination step.
In Europe and the Asiatic regions, the KAPA SYBR Fast qPCR Master Mix can be ordered from Millipore Sigma. In Canada, depending upon the territory, it may be available from either Roche or Millipore Sigma.
We have done limited testing with alternative SYBR qPCR mixes which in most cases did not yield comparable results to KAPA. In cases that customers do not have access to the KAPA SYBR Fast qPCR Master Mix due to product backorder and cannot avoid using an alternative qPCR Master Mix, please reach out to 10x Genomics Technical Support (firstname.lastname@example.org) for guidance.
Note: Due to limited testing performed, 10x Genomics does not guarantee assay performance if an alternative qPCR master mix is used.
Products: Visium for fresh-frozen, Visium for FFPE