Name: James Beauchamp Open PCR Machine Engineer(s)
Name: Student Role(s)
Name: Student Role(s)
Name: Student Role(s)
Name: Student Role(s)
LAB 2 WRITE-UP
Thermal Cycler Engineering
Our re-design is based upon the Open PCR system originally designed by Josh Perfetto and Tito Jankowski.
System Design
Key Features
Instructions
Protocols
Materials
Supplied in Kit
Amount
10 μM Forward Primer
2.5 μL
10 μM Reverse Primer
2.5 μL
GoTaq master mix
119.5 μL
dH2O
119.5 μL
Total Volume
250 μL
Supplied by User
Amount
DNA Sample (20ng)
.5 μL
PCR Protocol
Plug PCR machine into the computer.
Open the 'OpenPCR' program on the computer.
Label the tubes. This information should include the patient number (1 or 2) as well as the replication number. The positive and negative control should also be labeled.
Prepare the experiment by loading the reactants into the PCR tubes. This will consist of the patients DNA, along with the master mix components. As each tube is filled, put it into the chamber at the top of the machine.
Close the lid of the chamber.
Customize the settings in the 'Thermal Cycler' program to include three stages. Stage 1 is one cyle and in it the reactancts will heat up to 95 degrees Celcius for three minutes. During Stage 2, there are 35 cycles. The reactancts will heat to 95 degrees Celsius for 30 seconds, 57 degrees Celsius for 30 seconds, and 72 degrees Celsius for 30 seconds.
Press start on the program to begin running the PCR.
BME 103 Fall 2012
