Notes from Donita and Donna (3-10-11):
PV92 PCR INSTRUCTION SUMMARY FOR BIO-RAD MyCycler THERMAL CYCLER
IMPORTANT: Safety of students is the teacher’s responsibility. Prior to using the thermal cycler in the classroom, please refer to the safety procedures in the manufacturer’s instruction manual at
_____ Select a level location with at least 4 inches clearance to the left, right, and rear.
_____Plug the power cord into a properly grounded outlet. A quick diagnostic routine will be
performed. Once this routine is completed, the Home screen is displayed.
_____ A green LED light comes on next to the “Standby” button. There is no “on” or “off”
switch or button.
_____From the Home screen, press F1 Protocol Library.
_____ Use the arrow keys to highlight pv92pcr. Press Enter.
_____Use the yellow down arrowkey to select View Protocol. Press Enter. This is a
good moment to show the students the Run Protocol. (Use arrow to view entire
screen.) Make sure the protocol is correct, but you should not have to change
anything. Press F5 when done.
_____Select Run Protocol from the selection box. Press Enter.
_____ On the RUN SETUP screen the MODE choice should be Algorithmic
Measurement.
_____On the RUN SETUP screen, the Thermal Cycler defaults to 50 µL for the Sample
Volume. Use the yellow arrow buttons to move to the Sample Volume. Change to
40 µL. Hit Enter.
_____ On the RUN SETUP screen it will ask you if you want a hot start temperature.
Select “NO”. Hit Enter.
_____Place PCRtubescontaining the three controls and the students’ DNA
into the green plastic sample loading tray. Make sure that there are at least 10
empty tubes around the perimeter of the sample loading tray. This keeps
the lid from smashing your tubes during the course of the protocol. Close lid.
_____Press F5-Begin Run. Protocol execution will begin as soon as the heated lid reaches
the appropriate temperature. The amplification process will take almost 3 hours to complete. However, the machine can be left running overnight. When the process is complete, the machine will refrigerate the samples until you retrieve them the next day.
_____ The End of Run screen is automatically displayed after protocol execution. Hit Stop and
then End Run.