BME100 f2013:W1200 Group14 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: student Role(s) Name: student Role(s) Name: student Role(s) Name: student Role(s) Name: student 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 9 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 ... (did what? fill in your answer) When we unplugged the white wire that connects (part 6) to (part 2), the machine ... (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: CT Patient 1 ID: 30337 Replicate 1 Tube Label: 303 Patient 1 ID: 30337 Replicate 2 Tube Label: 304 Patient 1 ID: 30337 Replicate 3 Tube Label: 305 Negative Control: Non-Cancer DNA Template, Tube Label: NCT Patient 2 ID: 57707 Replicate 1 Tube Label: 577 Patient 2 ID: 57707 Replicate 2 Tube Label: 578 Patient 2 ID: 57707 Replicate 3 Tube Label: 579 Materials PCR Reaction mix, 8 tubes (50 micrometers), DNA primer mix, disposable pipette DNA Sample Set-up Procedure 1) Label all different tubes so we don't mix up any of the results. 2) Add 25μL of DNA sample mix to each corresponding tube, using different disposable pipette tips for each tube. 3) Place the tubes into a thermocycler 4) Program the Thermocycler to run as follows so PCR occurs. Thermocycler Program 1. 1 Cycle at 95°C for 3 minutes 2. 35 Cycles at 95°C for 30 seconds, 57°C for 30 seconds, 72°C for 30 seconds 3. 72°C for 3 minutes 4. Hold the mix at 4°C PCR Reaction Mix The PCR Reaction mix has Taq DNA Polymerase, MgCL2, and dNTP's. DNA/ primer mix The DNA primer mix has the same forward primer and reverse primer for all tubes Research and Development PCR - The Underlying Technology (Add a write-up, essay-style, organized into paragrpahs with descriptive headers, based on the Q&A's from Section three of your worksheet) PCR, or polymerase chain reaction, is used to amplify DNA to diagnose disease, map the human genome, or even to solve crimes. There are several components involved in completing PCR-- the template DNA, the primers, the Taq polymerase, magnesium chloride, and deoxyribonucleotides. The purpose of each component is as follows: Template DNA: T Primers: Taq polymerase: Magnesium chloride: Deoxyribonucleotides: (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.)