OUR TEAM

Name: Benjamin Hook Protocol

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

PCR Protocol
Step 1: Download the Open PCR Software onto Computer
Step 2: Plug in and turn on the Open PCR machine, connect the USB cable to your computer
Step 3: On the machine interface, select "DNA replication" and then choose desired number of cycles (at least 30 for good results).
Step 4: Using the Pipette, transfer 32 samples of the patients DNA into each test tube. You should only use 1 Pipette tip for this part of the process.
Step 5: Next, transfer the forward and reverse primers into each of the 32 test tubes. 2 Pipette tips should be used in this part of the process: 1 for all of the forward primer transfers, and 1 for all of the reverse primer transfers.
Step 6: Using a new pipette tip, transfer the GoTaq Master mix into each of the 32 test tubes.
Step 7: Dilute the 32 solutions by filling the remainder of the test tube with distilled water.
Step 8: Carefully Label Each test tube with a sharpie making sure that the positive and negative controls are clearly marked.
Step 9: Open the heated lid of the Open PCR machine and place the test tubes into the designated slots. Close the lid.
Step 10: Make sure everything is properly connected and then choose "begin replication" on the interface.
Step 11: Check to make sure that the computer is correctly receiving the information, if not: stop the cycle, check the USB cord, and try again.

DNA Measurement Protocol
Step 1: When replicating is finished, remove the 32 tubes from the PCR Machine.
Step 2: Label the pipettes to match up with the correlating test tubes.
Step 3: Use one pipette for the SYBR Green and one pipette for waste.
Step 4:

Research and Development

Background on Disease Markers

Primer Design

Illustration