Ah, the classic three-step PCR dance. In our FSL, denaturation is at 94–95°C to split the DNA strands, annealing at 50–65°C for primers to bind, and extension at 72°C for the Taq polymerase to work. But yaar, our old thermal cycler is so temperamental, we often run the annealing a degree or two lower than the manual says—it just works better with our dusty samples from the mofussil. You learn the real protocol from the machine’s moods, not just the textbook.
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