BME103:W930 Group2: Difference between revisions
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How PCR Works:<br> | How PCR Works:<br> | ||
Step 1: Denaturation by Heat <br> | Step 1: Denaturation by Heat <br> | ||
Initially, heat separates a DNA strand into two separate strands. This is allowed because the hydrogen bonds holding the DNA together are weak and easily separable when heated. <br> | Initially, heat separates a DNA strand into two separate strands. This is allowed because the hydrogen bonds holding the DNA together are weak and easily separable when heated. <br> | ||
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After the primers bind to the complementary DNA, the temperature is raised and the enzyme Taq DNA polymerase is used to replicate the strands. The Taq DNA polymerase, active at high temperatures, facilitates the binding and joining of the complementary nucleotides. It synthesizes an identical double-stranded DNA strand. Extension begins at the 3’ end of the primer because Taq DNA polymerase synthesizes exclusively in the 5’ to 3’ direction, so the free nucleotides are only added to the 3’ end of the primer. | After the primers bind to the complementary DNA, the temperature is raised and the enzyme Taq DNA polymerase is used to replicate the strands. The Taq DNA polymerase, active at high temperatures, facilitates the binding and joining of the complementary nucleotides. It synthesizes an identical double-stranded DNA strand. Extension begins at the 3’ end of the primer because Taq DNA polymerase synthesizes exclusively in the 5’ to 3’ direction, so the free nucleotides are only added to the 3’ end of the primer. | ||
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How to Amplify a Patient’s DNA sample: <br> | How to Amplify a Patient’s DNA sample: <br> | ||
1. A PCR Machine is created to amplify a patient’s DNA sample. In order to use a PCR machine, the steps include to first gather the DNA samples, 50 μL each. | 1. A PCR Machine is created to amplify a patient’s DNA sample. In order to use a PCR machine, the steps include to first gather the DNA samples, 50 μL each. | ||
2. The frozen DNA samples are separated into tubes and once melted, 8 transfer pipettes were used to add primer. | 2. The frozen DNA samples are separated into tubes and once melted, 8 transfer pipettes were used to add primer. |
Revision as of 10:44, 14 November 2012
BME 103 Fall 2012 | 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 Testing
When we unplugged part mounting plate from circuit board, the machine had no visible screen and we could not see the the information on the LCD screen. When we unplugged the white wire that connects the circuit board to the sample holder, the machine's temperature was unable to be monitored and could not be regulated
Our group did our test run on October 24, 2012. During the test run, we ran into no problems with the machine. We were done and found our results in a matter of 1 to 2 hours. The Open PCR machine was connected to one computers to control each temperature of each cycle of PCR during the experimental.
ProtocolsPolymerase Chain Reaction How PCR Works: Step 1: Denaturation by Heat
1. A PCR Machine is created to amplify a patient’s DNA sample. In order to use a PCR machine, the steps include to first gather the DNA samples, 50 μL each. 2. The frozen DNA samples are separated into tubes and once melted, 8 transfer pipettes were used to add primer. 3. Once the samples are ready, we processed the DNA in the openPCR system.
400μM dATP, 400μM dGTP, 400μM dCTP, 400μM dTTP and 3mM MgCl2, reaction buffer (pH 8.5), nuclease-free water
Flourimeter Measurements
Fluorimeter assembly procedure:
How to open images in Image J: Research and DevelopmentSpecific Cancer Marker Detection - The Underlying Technology Cancer Associated with the gene: Breast and Colorectal cancer with a susceptibility to LiFraumeni Syndrome Source: http://omim.org/entry/604373?search=CHEK2&highlight=chek2 Normal Gene Sequence: Cancer Gene Sequence: Baye's Rule:
ResultsDescription--------INTDEN Subtracted Background----------DNA Concentration (micrograms/mL) Water Blank----------218527------------------------------- 0
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