Molecular biology master mix preparation question?

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    • #14887
      cryocell
      Participant

      okay guys so…how would I prepare this master mix…

      I need 48 μL master mix total which will be added to 2 μL of template to create a 50μL total reaction volume…

      how much …
      5x taq buffer would i need @ a 1x concentration
      taq DNA pol 5U/μL at a concentration of 1.25 U would I need?
      Forward Primer 10 μM at 0.2 μM concentration?
      Reverse Primer 10 μM at 0.2μM concentration?
      and how much nuclease free water would be added?

      also 1 μL of nucleotide 10 mM mix is added at a concentration of 0.2 mM to the master mix.

    • #104694
      JackBean
      Participant

      is that really so hard to calculate? 🙄

      If you’re pipetting it one solution at a time, than it’s no master mix. Master mix is done for example in amount of 1 ml and then aliqouted.

      The final volume is 50 ul and you need to
      5x dilute buffer
      5/1.25-times dilute Pol
      10/0.2-times dilute both primers
      10/0.2-times dilute the nucleotide mix (which is 1 ul, so that should tell you something about the primers;)
      and add water up to 48 ul

    • #104705
      stephroche
      Participant

      Buffer 10µL
      Primers : 1µL each
      dNTP : 1µL
      Taq Pol : 0.25µL
      H2O : 34.75µL

      + 2µL DNA

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