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==PCR== *Dilute SYBR green I to 20X, then add 0.2X to PCR tube *Run PCR tube in BioRad DNAEngine by using Opticon Monitor 3 program *If no automatic connection, click on "Instrument" --> "Scan for instrument" *If no "prepare new run" under "Master", click on "open" --> open an old file, then edit *When editing the plate setup, choose SBG1 for SYBR green I detector *When editing protocol setup, add "plate read" before repeating the cycle *When hit run, it automatically asks for saving the running file *After a few running cycles, check "Status" and "Quantitation" to visualize PCR process *20 cycles need more than 3hrs to finish *No saturation was observed in any samples after 20 cycles, even for 500pg *NTC showed similar amplification as other samples, with a broader range of curves
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