BME100 f2013:W900 Group15 L4

BME 100 Fall 2013	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: Justin Dombrowski Protocol Planning Name: Saiswathi Javangula Researcher and Developer Name: Gage Bebak Open PCR Machine Testing Name: Ryan Fisher Protocol Planning Name: Abdulrahman Researcher and Developer Name: student Role(s) LAB 1 WRITE-UP Initial Machine Testing The Original Design (Add image of the full OpenPCR machine here, from the Week 9 exercise. Write a paragraph description for visitors who have no idea what this is) Experimenting With the Connections When we unplugged (part 3) from (part 6), the machine ... (did what? fill in your answer) When we unplugged the white wire that connects (part 6) to (part 2), the machine ... (did what? fill in your answer) Test Run (Write the date you first tested Open PCR and your experience(s) with the machine) Protocols Thermal Cycler Program DNA Sample Set-up row 1 cell 1 row 1 cell 2 row 1 cell 3 row 1 cell 4 row 2 cell 1 row 2 cell 2 row 2 cell 3 row 2 cell 4 DNA Sample Set-up Procedure Step 1 Step 2 Step 3... PCR Reaction Mix What is in the PCR reaction mix? DNA/ primer mix What is in the DNA/ primer mix? Research and Development PCR - The Underlying Technology Function of each component in PCR reaction"' The function of the template DNA during PCR reaction is to create a complimentary strand to result in one DNA strand that will convert into two. Primers are used when one attaches to one end of the segment and the primer attaches to the bottom strand of the other end. The Taq polymerase copies a cell's DNA before it divides in two. Magnesium chloride is important to the reaction as it is added to allow the presence of magnesium into the reaction as it acts a catalyst as the reaction cannot proceed without magnesium. The deoxyribonucleotides grab nucleotides that are floating in the liquid around it and attaches them to the end of the primer. Steps of thermal cycling During the initial step of 95 degrees Celsius for 3 minutes, the DNA separates from its double helix formation, allowing the two strands of DNA to be parallel to one another. During the step of Denature at 95 degrees Celsius for 30 seconds, the double stranded DNA separates or denatures into single strands. Next, is the process of anneal at 57 degrees Celsius for 30 seconds, the primers bind or anneal to complimentary matches on the target DNA sequence. During the next step of extend, the enzyme Taq polymerase binds to the prime sequences and adds nucleotides to extend the second strand. During the final step at 72 degrees celsius for 3 minutes, the target sequence defined by the primers begins to accumulate. During the final hold of four degrees Celsius, the remaining DNA left in the sample after the cycling is identified. During the base-pairing of the bonding, Adenine (A) sticks to Thymine (T), while Cytosine (C) sticks to Guanine (G). (BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the PCR video/ tutorial might be useful. Be sure to credit the sources if you borrow images.)