PCR Using Q5® Hot Start High-Fidelity DNA Polymerase (M0493). 
Set up the following reaction:
Component25 µl Reaction50 µl ReactionFinal Concentration5X Q5 Reaction Buffer5 µl10 µl1X10 mM dNTPs0.5 µl1 µl200 µM10 µM Forward Primer1.25 µl2.5 µl0.5 µM10 µM Reverse Primer1.25 µl2.5 µl0.5 µMTemplate DNAvariablevariable< 1,000 ngQ5 Hot Start High-Fidelity DNA Polymerase0.25 µl0.5 µl0.02 U/µl5X Q5 High GC Enhancer (optional)(5 µl)(10 µl)(1X)Nuclease-Free Waterto 25 µlto 50 µl.  
Gently mix the reaction.
Collect all liquid to the bottom of the tube by a quick spin if necessary and overlay the sample with mineral oil if using a PCR machine without a heated lid.
Transfer PCR tubes to a PCR machine and begin thermocycling.
