■ PAUSE POINT Purified PCR products can be stored at –20 °C for up to several months. 79DNA heteroduplex formation. Set up the annealing reaction as follows: ComponentAmount (μl)Taq PCR buffer, 10×2Normalized PCR product, 20 ng μl−118Total volume2080Anneal the reaction by using the following conditions: Cycle numberCondition195 °C, 10 min295-85 °C, −2 °C s−1385 °C, 1 min485-75 °C, −0.3 °C s−1575 °C, 1 min675-65 °C, −0.3 °C s−1765 °C, 1 min865-55 °C, −0.3 °C s−1955 °C, 1 min1055-45 °C, −0.3 °C s−11145 °C, 1 min1245-35 °C, −0.3 °C s−11335 °C, 1 min1435-25 °C, −0.3 °C s−11525 °C, 1 min1625-4 °C, −0.3 °C s−1174 °C, hold81SURVEYOR nuclease S digestion. Master-mix and add the following components on ice to the annealed heteroduplexes from Step 80, to a final volume of 25 μl: ComponentAmount (μl)Final concentrationAnnealed heteroduplex20MgCl2 stock solution supplied with kit, 0.15 M2.515 mMddH2O0.5SURVEYOR nuclease S11×SURVEYOR enhancer S11×Total25
