BME100 s2018:Group9 W0800 L4

BME 100 Spring 2018	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: Anna Avila Name: Taylor Dinubilo Name: Tyler Thompson Name: Zoey Wirtes Name: student Role(s) Name: student Role(s) LAB 4 WRITE-UP Protocol Materials Lab coat and disposable gloves PCR reacon, mix, 8 tubes, 50 μL each: Mix contains Taq DNA polymerase, MgCl2, and dNTP’s (hp://www.promega.com/resources/protocols/product‐informaon‐sheets/g/gotaq‐colorless ‐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 pipee ps: only use each only once. Never reuse disposable pipee ps. If you do, the samples will become cross‐contaminated Cup for discarded ps Micropipeor OpenPCR machine: shared by two groups PCR Reaction Sample List Tube Label PCR Reaction Sample Patient ID G9 + Positive control none G9 - Negative control none G9 1-1 Patient 1, replicate 1 44954 G9 1-2 Patient 1, replicate 2 44954 G9 1-3 Patient 1, replicate 3 44954 G9 2-1 Patient 2, replicate 1 75503 G9 2-2 Patient 2, replicate 2 75503 G9 2-3 Patient 2, replicate 3 75503 DNA Sample Set-up Procedure Step 1: Extract DNA from cells. step 2: Place the DNA into a special PCR tube. Step 3: Add primer 1 to the PCR tube. Step 4: Add primer 2(will attach to the second site) Step 5: Add nucleotide to the PCR tube. Step 6: Add DNA Polymerase to the PCR tube. Step 7: Place PCR tube, with all components, into a DNA thermal cycler. OpenPCR program Heated lid: 100°C Initial step: 95°C for 2 minutes Number of Cycles: 25 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 Research and Development PCR - The Underlying Technology What is the function of each component of a PCR function? What happens to the components during each step of thermal cycling? Base-pairing. driven by hydrogen bonding, allows base pairs to stick together. Which base anneals to each listed? Adenine: Thymine: Cytosine: Guanine: During which two steps of thermal cycling does base-pairing occur? Explain. SNP Information & Primer Design Background: About the Disease SNP Primer Design and Testing