PCR with Q5® High - Fidelity 2X Master Mix ( M0492 )
Set up the following reaction on ice : Q5 High - Fidelity 2X Master Mix , 12.5 µl , 25 µl , 1X ; 10 µM , Forward Primer , 1.25 µl , 2.5 µl , 0.5 µM ; 10 µM , Reverse Primer , 1.25 µl , 2.5 µl , 0.5 µM ; Template DNA variable variable < 1,000 ng Nuclease - Free Water , to 25 µl , to 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 .
Quickly transfer PCR tubes to a thermocycler preheated to the denaturation temperature ( 98°C ) .
