BME103:T930 Group 11 l2

BME 103 Fall 2012	Home People Lab Write-Up 1 Lab Write-Up 2 Lab Write-Up 3 Course Logistics For Instructors Photos Wiki Editing Help
OUR TEAM Name: Benjamin Hook Protocol Name: Jacqueline Janssen Protocol 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 Open PCR Machine (32) Plastic Test Tubes to Fit into PCR Machine PCR Power Adapter USB Cable Flourimeter (8) Glass Slides Supplied by User Low Retention Adjustable Pipette with Disposable Pipette Tips DNA Solution Positive Control DNA Soltution Negative Control DNA Solution What the DNA Solution Should Consist Of 1 Micro-Liter Forward Primer 1 Micro-Liter Reverse Primer 50 Micro-Liter GoTaq Master Mix .2 Micro-Liter Patient's DNA (Or Controls) 47.8 Micro-Liter Distilled Water 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