18
CHAPTER 3 – Calibration
3.1 When to Calibrate the Open qPCR
The Open qPCR needs to be calibrated before its initial use. Calibration must also be run each
time any of the following events occur:
o
The lid height is adjusted
o
The tube type is changed
o
The machine is transported
3.2 Adjusting the Open qPCR Lid Height
When using the instrument for the first time, set the lid height for the 100
µ
L PCR tubes that will
be used in your qPCR runs. Calibrate the instrument for that specific lid height.
Figure 3.1
Open the lid by pressing the small silver button near the lid bottom. You should feel the latch
mechanism release.
There are 2 rows of 8 wells, accommodating either 8-strip 100
µ
L PCR tubes or individual 100
µ
L PCR tubes. Use gloves while handling the tubes as prints on the tubes may affect the
fluorescence measurements. Ensure there are 16 capped tubes in the heat block during the lid
calibration process.
Raise the lid height by turning the knob on the lid clockwise. Place the PCR tubes in the
instrument and then close the lid until it latches. Proceed to turn the knob counter-clockwise to
lower the lid. Set the knob so that it just touches the top of the tubes and you feel a light
resistance.
Test the lid height by opening the lid and making sure you can close it again easily with the
tubes in place. If it does not close easily and you need to adjust the height for the next set of
tubes, a recalibration of the instrument will be required.
Calibrate using the built-in procedure. If calibration fails, re-adjust the lid height and repeat the
calibration process.
