I know there have been some discussion about this before, but I want to bring it up again with a few specific concerns and see what people think.
Typically, for preamplification, I've been using NEB's Phusion Flex hot-start enzyme. It's high fidelity, and it gives good yields in our hands. Recently though, I'm starting to worry about chimeric products, and a few people have been telling me Phusion is not a good choice for that. They invariably reference to Q5 or Kapa HiFi.
There's a nature method paper (doi:10.1038/nmeth.1814) that also suggests that Kapa HiFi gives lower mutation rate, though looking around, some people seem to dispute this. I'm wondering if anyone else here have any input on this? I would of course hate to have to change to a new enzyme, but if there's a supposedly big difference...
Typically, for preamplification, I've been using NEB's Phusion Flex hot-start enzyme. It's high fidelity, and it gives good yields in our hands. Recently though, I'm starting to worry about chimeric products, and a few people have been telling me Phusion is not a good choice for that. They invariably reference to Q5 or Kapa HiFi.
There's a nature method paper (doi:10.1038/nmeth.1814) that also suggests that Kapa HiFi gives lower mutation rate, though looking around, some people seem to dispute this. I'm wondering if anyone else here have any input on this? I would of course hate to have to change to a new enzyme, but if there's a supposedly big difference...
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