BME100 s2015:Group3 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: Abraham Rosengard Error creating thumbnail: File missing Name: Juan Patterson Error creating thumbnail: File missing Name: Gabriella Leiva-Hidalgo Error creating thumbnail: File missing Name: Jessica Resendiz Error creating thumbnail: File missing Name: Raghad Tamim ] Error creating thumbnail: File missing Name: Your name 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 G3 + Positive control 91905 G3 - Negative control 11553 G3 1-1 Patient 1, replicate 1 G3 1-2 Patient 1, replicate 2 G3 1-3 Patient 1, replicate 3 G3 2-1 Patient 2, replicate 1 G3 2-2 Patient 2, replicate 2 G3 2-3 Patient 2, replicate 3 DNA Sample Set-up Procedure The materials and DNA samples of the two patients were obtained along with our positive and negative control samples. The strip of tubes containing the PCR mixtures was cut in half so two strips of four linked tubes were created. This step was done so the tubes fit into the PCR machine. The sides of the PCR mixture tubes were labeled by negative and positive control, along with three individual trials of both patients. This was done using the black marker. The tubes were placed into a rack. 50 micro liters of the negative control, positive control, and each patient were transported into individual PCR mixtures. Individual pipettes were disposed after each use. There was a slight deviation from the guidelines, and 100 ml of patient one was placed into the second trial single PCR mixture tube. 50 ml of this was then transported into the third trial of the first patient to dilute the second and third trials. The PCR tubes were then closed and placed into the PCR machine. After closing the lid, the PCR program was started. 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