BME103:T130 Group 9

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: Luke Lammers Protocol Planner Name: Bryce Munter R & D Scientist Name: Daniel Saman OpenPCR Machine Engineer Name: Adrian Munoz Protocol Planner Name: David Probst R & D Scientist LAB 1 WRITE-UP Initial Machine Testing The Original Design () Experimenting With the Connections When we unplugged the LCD from the Circut Board, the display was disconnected and would not appear.The PCR would still work. When we unplugged the white wire that connects the PCB Circut Board to Temperature Sensor, the machine would not be able to sense temperature correctly and would not give us a reading. Test Run (Write the date you first tested Open PCR and your experience(s) with the machine) Protocols Polymerase Chain Reaction Polymerase Chain Reaction (PCR) works by heating up a DNA sample in order to allow DNA primers to bind and replicate the desired gene; this process is repeated over and over again over a several hour duration in order to amplify the amount of that gene in the sample. This creates a testable amount of the DNA in order to screen it for certain cancer-associated polymorphisms, for instance. PCR involves heating the solution containing DNA in order to denature and unwind the DNA, separating the two. The mixture is then cooled slightly, allowing the DNA primers to bind to the single strands and begin coding the desired gene. This creates a second DNA sample that can then undergo PCR in addition to the original DNA sample. This process is repeated several times over several hours in order to get a large enough amount of the desired gene to test. The process of PCR involves three basic steps that are repeated many times: 1. Denaturation: 2. Annealing: 3. Elongation: Reagent Volume Template DNA (20 ng) 0.2 µL 10 µM forward primer 1.0 µL 10 µM reverse primer 1.0 µL GoTaq master mix 50.0 µL dH2O 47.8 µL Total Volume 100 µL Eight samples were tested during this investigation. These included: a positive control with the cancer DNA template, a negative control without the cancer DNA template, and three samples each from the two subjects. The first subject was a 46 year old woman, correlating to test tubes labeled 2, and the other was a 62 year old man, correlating to test tubes labeled 4. Flourimeter Measurements (Add your work from Week 3, Part 2 here) Research and Development Specific Cancer Marker Detection - The Underlying Technology (Add a write-up of the information discussed in Week 3's class) (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 OpenPCR tutorial might be useful. Be sure to credit the source if you borrow images.) Results (Your group will add the results of your Fluorimeter measurements from Week 4 here)