BME103:T930 Group 7: Difference between revisions
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(Add a write-up of the information discussed in Week 3's class)<br> | (Add a write-up of the information discussed in Week 3's class)<br> | ||
PCR, or Polymerase Chain Reaction, is a process used to make copies of the same DNA sequences. This process includes a template DNA strand, which serves as the DNA that will be replicated. Primers are also needed to artificially synthesize the DNA strand. Taq polymerase then matches base-pairs, thus replicating the DNA. Magnesium Chloride is also needed because it binds to Taq, allowing it to function. Finally, dNTP’s are the nucleotides, A, | PCR, or Polymerase Chain Reaction, is a process used to make copies of the same DNA sequences. This process includes a template DNA strand, which serves as the DNA that will be replicated. Primers are also needed to artificially synthesize the DNA strand. Taq polymerase then matches base-pairs, thus replicating the DNA. Magnesium Chloride is also needed because it binds to Taq, allowing it to function. Finally, dNTP’s are the nucleotides, A, T, C, and G, that are used to make the new DNA. | ||
The process includes the following: | The process includes the following: <br> | ||
1. Heat to 95 degrees C. This separates the complementary DNA strands | 1. Heat to 95 degrees C. This separates the complementary DNA strands <br> | ||
2. Cool to 57 degrees C. This allows the primers to bind to the DNA strand. | 2. Cool to 57 degrees C. This allows the primers to bind to the DNA strand.<br> | ||
3. Heat to 72 degrees C. This allows Taq to extend the DNA copy | 3. Heat to 72 degrees C. This allows Taq to extend the DNA copy<br> | ||
4. Repeat | 4. Repeat | ||
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Normal Sequence: AAACTCTTACA<font color="red">'''T'''</font>TGCATACA <br> | Normal Sequence: AAACTCTTACA<font color="red">'''T'''</font>TGCATACA <br> | ||
the red C, specifically, makes the gene cancerous | the red C, specifically, makes the gene cancerous | ||
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Bayes' rule campares the odds of one event to another. We use it to predict the reliability of the PCR. | Bayes' rule campares the odds of one event to another. We use it to predict the reliability of the PCR. |
Revision as of 16:12, 13 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 TestingThe Original Design Description The OpenPCR Machine creates many copies of a small strand of DNA. In order to duplicate these DNA strands the PCR Machine must use many different temperatures during denaturing, annealing and extension. Denaturing is where the two strands of DNA separate. Annealing is where the DNA primer binds to the separate strands. Extension is when Taq Polymerase copies the DNA strands.
Experimenting With the Connections
When we unplugged the white wire that connects the OpenPCR circuit board to the main heating block, the temperature reading on the LCD screen changed.
The date the machine was used was on Thursday October 24th, 2012 10:32:32. The team's experience with the device was as follows:
Lightweight Silent User Friendly Great Software (it was very easy to use)
Took too long to complete its task Needed a computer Hard open the lid Not Aesthetically Pleasing Flammable (Wood + Extreme Heat=A Bad Situation Waiting to Happen.)
ProtocolsPolymerase Chain Reaction 1. Heat Denaturation: The heating of DNA to 95 degrees celsius allowed for the separation of the two strands of DNA. The nucleotides lose their base pair partners as the DNA is separated into a positive and a negative strand. 2. Annealing: The DNA now undergoes cooling of 57 degrees celsius to assist the process of annealing. Two primers are necessary for DNA replication as it's the primers that identify the specific targeted strand of DNA. Binding to the complementary sequence, the primers begin to produce the replication that's desired. 3. Extension: To finish off the first cycle of PCR, the temperature is once again raised to 72 degrees celsius. The enzyme Taq DNA polymerase then creates the new DNA strands by making each single strand now a double strand using the complementary sequences produced in annealing. The conclusion of these three steps is the production of two new DNA strands that are the replicate of the original strand.
PRC Master Mix Components: - Bacterially derived Taq DNA polymerase - dNTPs - Magnesium Chloride - Reaction buffers
(Add your work from Week 3, Part 2 here)
Research and DevelopmentSpecific Cancer Marker Detection - The Underlying Technology (Add a write-up of the information discussed in Week 3's class)
A SNP, or single nucleotide polymorphism, is a point mutation, meaning one nucleotide in a sequence is changed. This change is stable over many generations, and is present in at least 1% of the population. This makes the SNP ideal for a gene marker. The following SNP sequence was used as a primer for the cancerous gene
Results
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