BME100 s2015:Group18 12pmL4

BME 100 Spring 2015	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 Error creating thumbnail: File missing Name: Walter C. Bregon Role(s): Chief of Neurosurgery Error creating thumbnail: File missing Name: Jenna Taras Role(s) Error creating thumbnail: File missing Name: Eyerusalem Role(s) Error creating thumbnail: File missing Name: Nathan LeFort Role(s): being superior Error creating thumbnail: File missing Name: Abdurrahman Darwish Role(s) Error creating thumbnail: File missing Name: Hau Nguyen Role(s):Research & Development LAB 4 WRITE-UP Protocol Materials Lab coat and disposable gloves PCR reaction mix, 8 tubes, 50 μL each: Mix contains Taq DNA polymerase, MgCl2, and dNTP's (http://www.promega.com/resources/protocols/product-information-sheets/g/gotaqcolorless-master-mix-m714-protocol/) DNA/primer mix, 8 tubes, 50 μL each: Each mix contains a different template DNA. All tubes have the same forward primer and reverse primer A strip of empty PCR tubes Disposable pipette tips: only use each only once. Never re-use disposable pipette tips or samples will be cross-contaminated Cup for discarded tips Micropipettor OpenPCR machine: shared by two groups PCR Reaction Sample List Tube Label PCR Reaction Sample Patient ID G18 P Positive control none G18 N Negative control none G18 1-1 Patient 1, replicate 1 41180 G18 1-2 Patient 1, replicate 2 41180 G18 1-3 Patient 1, replicate 3 41180 G18 2-1 Patient 2, replicate 1 68784 G18 2-2 Patient 2, replicate 2 68784 G18 2-3 Patient 2, replicate 3 68784 DNA Sample Set-up Procedure Collect a small sample of DNA from cells such as blood, skin, saliva or even hair follicles Move your extracted DNA sample into a special PCR tube Add primer 1 to the PCR tube (Primer 1 attaches to a site on the DNA strand at either end of the template segment) Add primer 2 (which attaches to the 2nd site of the template segment on the DNA strand) to the PCR tube Add nucleotides to the PCR tube Add DNA polymerase to the PCR tube Now place the PCR tube containing all of the reaction components into the DNA thermal cycler Push start button on DNA thermal cycler OpenPCR program HEATED LID: 100°C INITIAL STEP: 95°C for 2 minutes NUMBER OF CYCLES: 35 Denature at 95°C for 30 seconds, Anneal at 57°C for 30 seconds, and Extend at 72°C for 30 seconds FINAL STEP: 72°C for 2 minutes FINAL HOLD: 4°C Research and Development PCR - The Underlying Technology Q1. What is the function of each component of a PCR reaction? Q2. What happens to the components (listed above) during each step of thermal cooling? Q3. DNA is made up of four types of molecules called nucleotides, designated as A, T, C and G. Base-pairing, driven by hydrogen bonding, allows base pairs to stick together. Which base anneals to each base listed below? Adenine-Thymine, Thymine-Adenine, Cytosine-Guanine, Guanine-Cytosine Bonus Round