BME103 s2013:T900 Group3

BME 103 Spring 2013	Home People Lab Write-Up 1 Lab Write-Up 2 Lab Write-Up 3 Course Logistics For Instructors Photos Wiki Editing Help
OUR TEAM Name: student Role(s) Name: student Role(s) Name: student Role(s) Name: student Role(s) Name: student Role(s) 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 Specific Cancer Marker Detection - The Underlying Technology Polymerase chain reaction is the process of copying DNA. Scientists may use this process to copy and study known cancerous DNA so that they may find resolutions to gene mutations. Deoxyribonucleic acid (DNA) is comprised of sugar phosphate chains and four main nucleotides which are guanine, adenine (purines), cytosine and thymine (pyrimidines). A single strand of DNA looks like a twisted rope ladder. The sugar-phosphate chains are like the side ropes of the rope ladder, and the four nucleotides are like the center ropes that extend perpendicularly from one side rope to the other and these “center ropes” are called rungs. Each rung is made-up of two different nucleotides. The nucleotides are base paired so adenine must always be paired with thymine and guanine must always be paired with cytosine. If they are not paired with their respective other, this would be called a mutation and could be cancerous. The idea is to obtain DNA with a gene mutation from a patient and amplify their DNA so further testing can be done to study, identify or to cure cancer. To perform the PCR, DNA is obtained, then a specific sequence of nucleotides is chosen and identified to be copied such as the gene mutation and this will be the template DNA. A forward and reverse primer is ordered to complement the template DNA. Adenine only attaches to thymine and cytosine only attaches to guanine. The DNA is mixed with the primers, dNTPs, Taq DNA polymerase, and magnesium chloride. The mix is heated to 95 degrees Celsius for 30 seconds which unzips the double helix strand in half between the base paired nucleotides. Then it is cooled to 57 degrees Celsius for 30 seconds which causes primer anneal binding on the ends of the nucleotides. It is heated to 72 degrees Celsius for 30 seconds, this causes the Taq polymerase which is catalyzed by the magnesium chloride to attach the buoyant dNTP’s to their complementary base pair. This completes one cycle and produces one copy. Performing this procedure 30 times would give approximately 1 million copies of the template DNA. (Add a write-up of the information discussed in Week 3's class) (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 OpenPCR tutorial might be useful. Be sure to credit the source if you borrow images.)