BME100 s2015:Group8 9amL4

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 Name: An Tran Role(s) Name: Molly Golek Role(s) Name: Anthony Gambirazio Role(s) Name: Austin Lehew Role(s) Name: Robert Culibrk Role(s) LAB 4 WRITE-UP Protocol Materials Lab Coat Disposable Gloves PCR Reaction Mix, 8 tubes, 50 uL each: Mix contatins Taq DNA polymerase, MgCL2, and dNTP's DNA/primer mix, 8 tubes, 50 uL 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 machines: shared by two groups PCR Reaction Sample List Tube Label PCR Reaction Sample Patient ID G8 P Positive control none G8 N Negative control none G8 1-1 Patient 1, replicate 1 G8 1-2 Patient 1, replicate 2 G8 1-3 Patient 1, replicate 3 G8 2-1 Patient 2, replicate 1 G8 2-2 Patient 2, replicate 2 G8 2-3 Patient 2, replicate 3 DNA Sample Set-up Procedure 1. Acquire the DNA samples from the patients as well as PCR reaction mix 2. Label all of the PCR tubes for all of the samples that will be tested 3. Place a new pipette tip on the micropipette 4. Take 50 microliters of the PCR reaction mix 5. Transfer PCR reaction mix to the corresponding labeled empty PCR tube 6. Discard the used pipette tip 7. Put a new pipette tip on the micropipette 8. Take 50 micro liters of patient DNA sample/primer corresponding to the label on the tube 9. Transfer the patient DNA into the tube containing the PCR reaction mix 10. Repeat steps 3 through 9 for all samples. Be sure to place the DNA in the correct labeled tubes 11. Place the tubes in the 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 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