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I have recently been told that the components of a PCR mixture need to be added in a specific order: primers, dNTP, and finally polymerase. May I know why this is the case?

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  • $\begingroup$ Welcome to the Biology Stack. Please take a tour and visit the help center for more information on what we do here. Please also note the homework policy, which requires questions to contain evidence that the asker has attempted to answer the question for themselves. On that note; what do you know about those components and why might you add polymerase last (what does it do, and what could it do if you add it earlier?)? $\endgroup$
    – bob1
    Commented Mar 26, 2023 at 19:50
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    $\begingroup$ you may be able to find your awnser here: biology.stackexchange.com/questions/79024/… $\endgroup$
    – Olaf
    Commented Apr 3, 2023 at 11:22

1 Answer 1

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Polymerase last: One reson is that pipetting conditions are vastly differing by person, I.e. a lot of people don't keep the reactions on ice. When no hot-start polymerase is used, pipetting it in too early in a room temp. reaction mix might lead to unintended reactions.

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