PCR Protocol for OneTaq® DNA Polymerase ( M0480 )
Set up the following reaction on ice :
Component25 μl reaction50 μl reactionFinal Concentration5X OneTaq Standard Reaction Buffer * 5 µl10 μl1X10 mM dNTPs ( # N0447 ) 0.5 µl1 μl200 µM10 µM Forward Primer0.5 µl1 μl0.2 µM10 µM Reverse Primer0.5 µl1 μl0.2 µMOneTaq DNA Polymerase0.125 µl0.25 µl1.25 units / 50 µl PCR * * Template DNAvariablevariable < 1,000 ngNuclease - 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 .
Quickly transfer PCR tubes to a thermocycler preheated to the denaturation temperature ( 94°C ) and begin thermocycling
