BME100 f2013:W1200 Group13 L4

BME 100 Fall 2013	Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help
OUR TEAM Name: Samantha Brenna(s) Name: student Role(s) Name: student Role(s) Name: student Role(s) Name: student Role(s) Name: student Role(s) LAB 1 WRITE-UP Initial Machine Testing The Original Design The picture above shows what the OpenPCR machine looks like while running and connected to the computer. The machine case is made of wood panels which contains the actual hardware of the device. The smaller box located on the top is the lid of the OpenPCR machine. This contains a 16-tube PCR block which is where the test vials are placed in order to complete the test run. The top part of the lid has a heat plate that heats the vials and also seals the experiment. Inside of the main body of the OpenPCR machine there is a heatsink, processor, led screen, and copper wire connections. The processor has a circuit board that recieves the data from and computes the calculations. The heatsink dispersses the heat and the fan attached cools down the device in order to avoid overheating. The led screen displays the data specifically the numbers of tests run and temperature. The machine also has an USB port and power connection in order to power the device and connect it to the computer in order to run the tests. Experimenting With the Connections When we unplugged the LED screen (part 3) from the circuit board(part 6), the machine didn't display anything on the screen because this connection is what allows the power to connect to the led screen. When we unplugged the white wire that connects (part 6) to (part 2), the machine lost the ability to record data and to sense the heat change in the machine. Test Run Our OpenPCR machine passed the testing that we conducted in class. We began the testing at 12:50 pm, and at 1:40pm the machine had run 19 out of 35 tests. When we concluded the experiment the temperature according to the screen on the device was 95 degrees Celsius, and had an ETA of 48. The test ran very smoothly for the first time use and we didn't encounter problems with the machine not working properly. The device was plugged into the computer and then we opened the program in order to start the tests. Then after entering in the information given we began running the test. There were no issues with the program or the machine. ) Protocols Thermal Cycler Program Stage 1: 95°C for 3 minutes-Double helix unwinds into one long strand Stage 2: (35 cycles of each) 1. 95°C for 30 seconds-Double helix breaks apart into two strands 2. 52°C for 30 seconds-Primers attach to target sequence 3. 72°C for 30 seconds-DNA polymerase begins constructing new strand Stage 3: 4°C for 3 minutes-Keeps solution of pure taget sequence at stable temperature DNA Sample Set-up PC 1R1 1R2 1R3 NC 2R1 2R2 2R3 DNA Sample Set-up Procedure Step 1 Place each smaple in its respective pipette tips. That is the positive control, negative control, patient 1, and patient 2. Step 2 Place the 4 pipettes inside the OpenPCR machine. Step 3 Run the machine for 1 cycle and record results. Step 4 Repeat step 3 untill the desired number of replications is reached. PCR Reaction Mix What is in the PCR reaction mix? The PCR reaction , 50uL each. the mix contains Taq DNA polymerase, MGCl2, and dNTP's. DNA/ primer mix What is in the DNA/ primer mix? Each mix contains a differant template DNA all tubes have the same forward primer and reverse primer. Research and Development PCR - The Underlying Technology (Add a write-up, essay-style, organized into paragrpahs with descriptive headers, based on the Q&A's from Section three of your worksheet) (BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the PCR video/ tutorial might be useful. Be sure to credit the sources if you borrow images.)