BME103 s2013:T900 Group2: Difference between revisions
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| [[Image:BME103student.jpg|100px|thumb|William Scott<br> R&D Scientist]] | | [[Image:BME103student.jpg|100px|thumb|William Scott<br> R&D Scientist]] | ||
| [[Image:BME103student.jpg|100px|thumb|Joe Sasnone<br> R&D Scientist]] | | [[Image:BME103student.jpg|100px|thumb|Joe Sasnone<br> R&D Scientist]] | ||
| [[Image: | | [[Image:404005_4622115081677_1186647743_n.jpg|100px|thumb|Mitch Riggs <br> Open PCR Machine Engineer]] | ||
| [[Image:BME103student.jpg|100px|thumb|Name: student<br>Role(s)]] | | [[Image:BME103student.jpg|100px|thumb|Name: student<br>Role(s)]] | ||
| [[Image:BME103student.jpg|100px|thumb|Name: student<br>Role(s)]] | | [[Image:BME103student.jpg|100px|thumb|Name: student<br>Role(s)]] | ||
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'''Experimenting With the Connections'''<br> | '''Experimenting With the Connections'''<br> | ||
When we unplugged (part 3) from (part 6), the | When we unplugged (part 3) from (part 6), the screen turned off. <br> | ||
When we unplugged the white wire that connects (part 6) to (part 2), the | When we unplugged the white wire that connects (part 6) to (part 2), the screen showed the wrong temperature. <br> | ||
'''Test Run''' | '''Test Run''' | ||
We first ran the machine on Tuesday, March 5. We used unit 3. We performed a simple test with the computer. The temperatures shown on the computer corresponded with the temperatures shown on the device indicating a successful test.<br> | |||
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# Step 3: Place the transfer pipette onto the pipette to prevent cross contamination (never re-use). | # Step 3: Place the transfer pipette onto the pipette to prevent cross contamination (never re-use). | ||
# Step 4: Use the pipette to transfer 50 microliters of each tube in the PCR reaction mix and transfer accordingly to the DNA sample tubes corresponding to the labels. | # Step 4: Use the pipette to transfer 50 microliters of each tube in the PCR reaction mix and transfer accordingly to the DNA sample tubes corresponding to the labels. | ||
# Step 5: Place the set of mixed tubes into the Open PCR machine and shut it tightly. | |||
# Step 6: Hook up the machine to a computer and run the Open PCR application with the pre set-up Thermal Cycler program. | |||
# Step 7: Once the application is done running, take out the tubes and hand it to the professor. | |||
'''PCR Reaction Mix''' | '''PCR Reaction Mix''' | ||
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'''DNA/ primer mix''' | '''DNA/ primer mix''' | ||
* The DNA/primer mix consists of samples of DNA of various patients. | * The DNA/primer mix consists of samples of DNA of various patients. DNA/ primer mix contains 8 tubes, 50 μL each: Each mix contains a different template DNA. All tubes have the same forward primer and reverse primer. | ||
Latest revision as of 11:24, 26 March 2013
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 TEAMLAB 1 WRITE-UPInitial Machine TestingThe Original Design Experimenting With the Connections When we unplugged (part 3) from (part 6), the screen turned off. When we unplugged the white wire that connects (part 6) to (part 2), the screen showed the wrong temperature.
We first ran the machine on Tuesday, March 5. We used unit 3. We performed a simple test with the computer. The temperatures shown on the computer corresponded with the temperatures shown on the device indicating a successful test.
ProtocolsThermal Cycler Program DNA Sample Set-up
DNA Sample Set-up Procedure
PCR Reaction Mix
DNA/ primer mix
Research and DevelopmentSpecific Cancer Marker Detection - The Underlying Technology Polymerase Chain Reaction (PCR) is a scientific method that utilizes DNA Polymerase to create a complimentary base strand from a template strand of DNA. Triphosphate nucleotides align with open DNA strands and DNA polymerase works to link the complementary nucleotide bases together growing strands through both condensation and hydroysis reactions. One major issue with DNA polymerase is that DNA strands are anti parallel, and polymerase is only able to add nucleotides to the free 3'OH group hence it can only build new strands in the 5'-3' direction (Sadava 279). Therefore, to correct for this issue the presence of a primer is required so that polymerase can proceed with directing the new nucleotides in place. Through these mechanisms it is possible to target specific positions on the template DNA sequence that a scientist intends to amplify(PCR 1). When the PCR process is completed the targeted DNA sequence containing the single-nucleotide polymorphism (SNP) will have manufactured over a billion copies (amplicons). The SNP included in the amplicon for this experiment was denoted as rs17879961. This polymorphism is a variant of the CHEK2 gene which if present in a person's genome may increase the risk of developing breast cancer (Brennan et al 1795). This SNP is signified by a single base change from a Thymine (T) to a Cytosine (C) located on chromosome 22. With the PCR system, forward and reverse primers can be designed to target the cancer gene abnormality and amplify/multiply it. The normal DNA strands may multiply, but not as exponentially as the abnormal cancer DNA strands. |