BME103:T930 Group 7
(→Initial Machine Testing)
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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
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 denaturingand extension.
Revision as of 12:26, 13 November 2012
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LAB 1 WRITE-UP
Initial Machine Testing
The 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:
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.)
Polymerase 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
- Magnesium Chloride
- Reaction buffers
(Add your work from Week 3, Part 2 here)
Research and Development
Specific Cancer Marker Detection - The Underlying Technology
(Add a write-up of the information discussed in Week 3's class)
The following sequence was used as a primer for the cancerous gene