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.
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