BME103:T930 Group 11

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: Tony Nguyen Role(s) Name: student Role(s) Name: Kenze Caulfield Lvl 13 Wizard Name: student Role(s) Name: Ben Hook Role (s) Name: student Role(s) LAB 1 WRITE-UP Initial Machine Testing The Original Design (Add image of the full OpenPCR machine here, from the Week 3 exercise. Write a paragraph description for visitors who have no idea what this is) Experimenting With the Connections When we unplugged (part 3) from (part 6), the machine's display screen didn't turn on or show up. When we unplugged the white wire that connects (part 6) to (part 2), the machine's temperature reading was altered compared to the accurate values from the computer. Test Run (Write the date you first tested Open PCR and your experience(s) with the machine) October 25, 2012. The machine was accurate and precise but the cycling was slow. Protocols Polymerase Chain Reaction The Polymerase Chain Reaction works by attaching MgCl to taq enzyme in the solution and then pulling deoxy nucleotide diphosphates, binding them to the strand that is already present. This gives a base to the DNA that is already there. DNA amplification contains three important steps: heat denauturation,primer annealing, and primer extension. Step One: Heat Denauturation-Heat is applied to the DNA and the two strands separate. Step Two:Primer Annealing-With excess dNTPs, oligonucleotides are added. Steph three: Primer Extension-DNA polymerase is added and new strands of DNA are synthesized. The DNA strands combine with the nucleotides to form completed, complimentary strands of DNA. PCR master mix contains Magnesium Chloride, all four Nucleotides needed to create DNA, and reaction buffers. Reagent volume Template DNA(20 ng) .2uL 10 uM forward primer 1.0 uL 10 uM reverse primer 1.0 uL GoTaq master mix 50.0 uL DH2O 47.8 uL Total Volume 100.0 uL Patient one's ID number is 12123, male, and 61 years old. Patient two's ID is 21312, female, and 56 years old. 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 100.0 μL 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 =