BME100 f2013:W1200 Group2 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: Amber Bengson Role: Open PCR Machine Testing Name: Chandler Varone Role: Open PCR Machine Testing Name: Prycilla Jones Role: Protocol Planning Name: Emily Mei Role: Protocol Planning Name: Terri Bivins Role: Research and Development LAB 1 WRITE-UP The Original Design (Write a paragraph description for visitors who have no idea what PCR is) Experimenting With the Connections When we unplugged (part 3) from (part 6), the machine screen went blank. (did what? fill in your answer) When we unplugged the white wire that connects (part 6) to (part 2), the machine read -40 degrees Celsius on the screen. (did what? fill in your answer) Test Run (Write the date you first tested Open PCR and your experience(s) with the machine) Protocols Thermal Cycler Program DNA Sample Set-up Positive control: cancer DNA template Tube Label: cancer ID Patient 1: 43849 Replicate 1 Tube Label: 1A ID Patient 1: 43849 Replicate 2 Tube Label: 1B ID Patient 1: 43849 Replicate 3 Tube Label: 1C Negative control: cancer DNA template Tube Label: not cancer ID Patient 2: 80853 Replicate 1 Tube Label: 2A ID Patient 2: 80853 Replicate 2 Tube Label: 2B ID Patient 2: 80853 Replicate 3 Tube Label: 2C DNA Sample Set-up Procedure Materials: -PCR Reaction mix: 8 tubes, 50 μL each -DNA/primer mix: 8 tubes, 50 μL each -Disposable pipettes -Patient’s samples (patient 1: 43849, patient 2: 80853) Procedure: Step 1:Determine positive and negative control. The negative control will not have the cancer marker. The positive control has cancer marker. Step 2:Label Tubes. Positive control: cancer negative control: Not cancer Patient 1 43849 Patient 2 80853 1A 2A 1B 2B 1C 2C Step 3:Add DNA/primer mix to positive control. Step 4:Add PCR reaction mix to positive control. Step 5:Place sample in Open PCR(thermo cycler) Step 6:While in the Open PCR the sample will be heated up to 95℃ the DNA chain separates into two strands. The temperature then decreases to 50℃ and the primers attach to the separate strands at the target sequence. The polymerase attaches to the primer and begins to replicate the strand. Step 7:Repeat steps 3-5 for the remaining tubes. Step 8:Compare Patient 1 DNA replicates to those of the positive and negative controls. step 9:Compare Patient 2 DNA replicates to those of the positive and negative controls. PCR Reaction Mix What is in the PCR reaction mix? -Taq DNA Polmerase -MgCl2 -dNTP's DNA/ primer mix What is in the DNA/ primer mix? Research and Development PCR - The Underlying Technology PCR Reaction Components The components of the PCR Reaction include the template DNA, primers, Taq Polymerase, Magnesium Chloride(MgCl2), and Deoxyribonucleotides (dNTP's). Each component or reagent is added to the PCR tube before running them through the PCR device and software called "Open PCR". The template DNA is the source of genetic information in which an enzyme copies from. The Taq Polymerase is an enzyme and specific kind of DNA Polymerase that is thermostable. It reads the genetic information form the DNA template and checks for mutations before copying data. Thermal Cycling Thermal cycling is the process of cycling a PCR tube holding PCR reaction in specific variations of time and temperature. The PCR reaction is first exposed to high temperatures, it is lowered, and increases slightly in temperature. The changes in temperature triggers and activates specific enzymes and/or reagents and is done ... Initial Step Denature Anneal Extend Final Step Final Hold Nucleotides and Base-pairing (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.)