Editing 2X qPCR Mix Protocol (section)

== Running the sample ==

:1. Keep the master mix and your sample apart until immediately before running.
:2. Dilute a portion of your samples to have a DNA concentration of 0.1-2ng/μL.
:3. Place 10μL of the diluted samples in each well in the plate.
:4. Lightly put the optical film over the plate.
:5. Take your prepped plate and your 2X master mix (in a tube on ice) down to the core facility.
:6. Start up the machine.
:7. Set up the following program.
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*Detector = SYBR, Rox = Yes (but only if you added it)

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*95°C for 10 min

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*95°C for 20sec

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*60°C for 60 sec

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*Repeat the two-step cycle 45x

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*If you want to, you can do a melt curve to be sure that your primers are only amplifying one thing.
::**Ramp from 50.0°C to 95.0°C
::**Read every 0.2°C h
::**Hold 00:00:02 between reads
:8. Add 10μL of 2X mix to each well (using a multi pipettor for this makes it much easier).
:9. Quickly centrifuge the plate.
:10. Place it in the machine and Run the program (you will need to save it first):