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==Procedures order== Step1: Testing the pH value of NS buffer + ALS buffer mixture (10 uL + 10 uL). The acceptibe pH is between pH 7~8. Step2: UV both NS and ALS for 10 min for decontamination. Step3: Prepare 2 stips of PCR tubes and transfer 1 uL template (diluted gDNA or H2O) into each of tubes as exp. design. Step4: Add 1.5 uL ALS into the tube and mix by pipetting. Incubate at RT for 3 min. Step5: Add 1.5 uL NS into the tube and mix by pipetting. Transfer the reaction on cool rack. Step6: Add 16 uL phi29 master mix into each of tubes. Mix by pipetting. Cap and label the tube. Step7: Place the reaction in Bio-Rad realtime PCR machine and start the reaction program.
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