BME103:T130 Group 16

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 Brayden Gallimore Research & Design Specialist/Data Compiler and Analyzer Name: Adam M White Experimental Protocol Planner/DNA Measurement Operator:Smartphone Name: Heather Borgard Open PCR Machine Tester 1/ImageJ Software Processor Name: Ashley Guerrero Open PCR Machine Tester 2000/R and D Scientist:Open Wet Ware Name: Zonash Zainab Protocol Person: Sample Prep and Application LAB 1 WRITE-UP The Original Design () Experimenting With the Connections When the wire connecting the LCD screen to the circuit board is unplugged, the LCD will not show, but the machine will still function. The screen will not present any premature values or any details about the thermal cycler. When the white wire connecting the circuit board to the temperature sensor is unplugged, the temperature will not be changed correctly. This would cause problems in the process of DNA amplification and the primers would most likely be unable to attach to the active sites. Test Run We first tested the PCR machine on 10/25/12 The Polymerase Chain Reaction Machine we used is a quick and inexpensive way amplify small sections of DNA. The main function of the PCR machine is to act as a DNA Thermal Cycler to isolate a specific section of DNA from a strand that has an excessive amount of genetic information. A target area on a strand of DNA is amplified and identical copies are generated using changes in temperature. The machine quickly and precisely heats and cools the DNA segments at different times over the span a a little over an hour. In cylce one, the thermal cycler heats to 95 degrees Celsius. The DNA begins to separate into two single strands. The thermal cylcer then cools to 50 degrees Celsius. The primers that were added in the tubes before the experiment then bind the target sites before the single strands pair up again. When the thermal cycle heats again to 72 degrees Celsius, DNA polymerase finds a primer to add complementary nucleotide to the strand.In cycle two the temperature is raised again to separate the DNA strands. When the temperature is lowered, the primers attach again. and the same process is repeated. In cycle three, two strands that begin with primer one and end with primer two appear. At then end of cylce four, there are even more copies of this fragment. As the cylces continue more and more copies of the target sequence will be generated. The this is especially useful to genetic mapping and detecting bio-markers. The PCR Machine is composed of a heated lid that presses against the tubes of DNA to heat during the cycles and thermal block where the tubes of DNA are placed, a heat sink, and a fan to absorb heat and cool during the different cycles, and an lcd screen to read the temperature and cylces that the thermal cycler is going though. Protocols Polymerase Chain Reaction (Add your work from Week 3, Part 1 here) 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 Sample Integrated Density DNA μg/mL Conclusion PCR: Negative Control E6 F6 G6 PCR: Positive Control E7 F7 G7 PCR: Patient 1 ID #####, rep 1 E8 F8 G8 PCR: Patient 1 ID #####, rep 2 E9 F9 G9 PCR: Patient 1 ID #####, rep 3 E10 F10 G10 PCR: Patient 2 ID #####, rep 1 E11 F11 G11 PCR: Patient 2 ID #####, rep 2 E12 F12 G12 PCR: Patient 2 ID #####, rep 3 E13 F13 G13 KEY Sample = Integrated Density = DNA μg/mL = Conclusion =