BME103:T930 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
Group 16 Name: Omar Moreno Salinas, and Rachel Juetten Open PCR, ImageJ Software Processor, Data Compiler, and Analyzers Name: Maggie McClure, and Swetha Swanminathan Protocol Persons: Sample Prep & Application Name: Marianna Singh DNA Measurement Operator Name: Muawiya Ali Al-Khalidi R&D Scientest LAB 1 WRITE-UP Initial Machine Testing The Original Design Experimenting With the Connections When the heat sink is unplugged from the circuit board, the LCD screen is turned off. When we unplugged the white wire that connected the circuit board to the heating block the temperature reading on the LCD screen dropped drastically. Test Run We first tested open PCR on October 18, 2012. We learned how to take accurate temperatures using the open PCR machine. Using open PCR we were able to make a polymerase chain reaction. In order for this to occur, open PCR had to send the DNA through different sets of temperatures to heat it up to separate the strands and expose the bases, then cool it down for the primers to bind to the sequences, and also heat it back up to attain an extension of the copy of the new DNA. Which was conducted in an hour and thirty minutes. Protocols Polymerase Chain Reaction Reagent Volume Template DNA (20ng) 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 100.0 μL Here is the patient information: 92136 M 54 Years 62276 F 61 Years Flourimeter Measurements Research and Development Specific Cancer Marker Detection - The Underlying Technology A Polymerase chain reaction is a machine that amplifies a single or a few strands of DNA to generate millions of copies of that DNA sequence. Using this technology scientists can determine whether a patient has a positive or negative result towards cancer. A method of getting this data is called the PCR detection method, a method that relies on thermal cycling, switching back and forth to melt DNA and then connect primers. This is a method that can be used to detect whether a patient has positive result for cancer, because a sample of DNA can be taken and whether that connects to the primers and creates a chain reaction, scientists can then determine whether this DNA is positive or negative towards cancer. An example of proving this method can be seen using the r17879961 SNP, a cancer-associated sequence, using the PCR detection method we can prove that r17879961 SNP is actually associated with cancer. Because it carries with the Polymerase chain reaction, and to further prove the patient has a positive result for cancer, we use fluorescent dye and if the DNA glows in the solution, then the results are positive for cancer. (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.) Insert non-formatted text here==Results== Sample Integrated Density DNA μg/mL Conclusion PCR: Negative Control 9880697 F6 G6 PCR: Positive Control 4981557 F7 G7 PCR: Patient 1 ID #####, rep 1 14455570 F8 G8 PCR: Patient 1 ID #####, rep 2 8591662 F9 G9 PCR: Patient 1 ID #####, rep 3 13212871 F10 G10 PCR: Patient 2 ID #####, rep 1 12472923 F11 G11 PCR: Patient 2 ID #####, rep 2 22091168 F12 G12 PCR: Patient 2 ID #####, rep 3 11477553 F13 G13 KEY Sample = Integrated Density = DNA μg/mL = Conclusion =