BME100 f2014:Group8 L4

BME 100 Fall 2014	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 Robert Schultz Isabella Germinario Bakir Mousa Daniel Hersey Ethan Lloyd Bailey Myers LAB 4 WRITE-UP LAB A 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/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 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 OpenPCR Program Settings The following values were used to run a heating & cooling program on the thermal cycler. 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 Part 1- The Function of Each Component of a PCR reaction- Template DNA- This provides a set of nucleotides for the primers to bond to and then to be replicated. Primers-These sets of nucleotides bind to the template DNA and begin the replication process. Taq Polymerase- This enzyme combines nucleotides to make a new strand of DNA which replicates the template strand Deoxyribonucleotides (dNTP's)- These are the building blocks for each DNA strand. These are combined to form new strands of the selected DNA. Part 2- The Steps of Thermal Cycling- Initial step: 95°C for 3 minutes- 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 3 minutes- Final hold: 4°C- Part 3- 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. Adenine anneals to Thymine Thymine anneals to Adenine Cytosine anneals to Guanine Guanine anneals to Cytosine Part 4- Base pairing occurs during the Annealing and Primer Extension phases LAB B