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Best in this case means: specific priming, long templates (amplifying whole plasmids >5 kb), and fast. I use NEB Q5 polymerase now which meets all of these requirements, but is very expensive...
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In my experiments, I've seen over 5 times now mCherry fusion proteins (N- and C-terminal) that work fine at ~23-30 C and show progressively less fluorescence at elevated temperatures during in...
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