BME100 s2015:Group16 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 Name: Hanna Monroe Name: Natkanes Pakjirasirikul Name: Trevaun Walker Name: Reaghan Fletcher Name:Thomas Murphy 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 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 G16 + Positive control none G16 - Negative control none G16 1-1 Patient 1, replicate 1 47360 G16 1-2 Patient 1, replicate 2 47360 G16 1-3 Patient 1, replicate 3 47360 G16 2-1 Patient 2, replicate 1 31303 G16 2-2 Patient 2, replicate 2 31303 G16 2-3 Patient 2, replicate 3 31303 DNA Sample Set-up Procedure Label Tubes with the labels from the table above (e.g. G16 +, G16 2-2...). Transfer the + and - controls to the PCR tubes labeled G16 + and G16 - using the micropipette. Transfer corresponding patient DNA to special PCR tube using the micropipette. Dispose and replace the tips between each transfer. Place the PCR reaction 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, 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 Question 1: Question 2: Question 3: Question 4: Base pairing occurs in the anneal step and final step of the thermal cycling. In the anneal step at 57°C for 30 seconds, the primer is pairing with the DNA strand. In the final step at 72°C for 3 minutes, the DNA polymerase is paring nucleotides to complete the DNA strand.