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==Step 1: PCR amplification reaction system setup== *Thaw Econo Taq on ice *Prepare strips and tubes on cooling rack *Add in Content in table to tubes accordinly {| border="1" align="center" | align="center" style="background:#f0f0f0;"|'''Content''' | align="center" style="background:#f0f0f0;"|'''Volume''' | align="center" style="background:#f0f0f0;"|'''X24''' | align="center" style="background:#f0f0f0;"|'''X48''' | align="center" style="background:#f0f0f0;"|'''X72''' | align="center" style="background:#f0f0f0;"|'''X96''' | align="center" style="background:#f0f0f0;"|'''1.5 mL Tube''' |- |Jan_09_CPG_D40809 (10uM)||align="right"|0.2uL||align="right"|4.8uL||align="right"|9.6uL||align="right"|14.4uL||align="right"|19.2uL||align="right"|2.4uL |- |Econo Taq||align="right"|50uL||align="right"|1250uL||align="right"|2400uL||align="right"|3600uL||align="right"|4800uL||align="right"|650uL |- |AP1V6 AP2V6 Mix (50uM)||align="right"|0.4uL||align="right"|9.6uL||align="right"|19.2uL||align="right"|28.8uL||align="right"|38.4uL||align="right"|4.8uL |- |RNAse free H2O||align="right"|50uL||align="right"|1200uL||align="right"|2400uL||align="right"|3600uL||align="right"|4800uL||align="right"|600uL |- |50X SYBG I||align="right"|0.4uL||align="right"|9.6uL||align="right"|19.2uL||align="right"|28.8uL||align="right"|38.4uL||align="right"|4.8uL |- |Total Volume|| align="right"| 101uL||align="right"|2424uL||align="right"|4848uL||align="right"|7272uL||align="right"|9696uL||align="right"|1212uL |} *Mix content in 1.5 mL tube vortex and spin down in the order of water>SYBR Green>Primer >template>econotaq *Aliquot 105uL using P200 pipette to strip tubes on cooling rack *Seal strips with strip caps *Place into Real Time PCR *Perform 94C 2M > (94C 30S > 58C 30S > 72C 30S>Plate Read) X16 > 72C 3M > 15C Hold
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