BME103:T130 Group 17
(→Research and Development)
(→Initial Machine Testing)
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the PCR and the machine <br>
Revision as of 18:11, 8 November 2012
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Lab Write-Up 1
Lab Write-Up 2
Lab Write-Up 3
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LAB 1 WRITE-UP
Initial Machine Testing
Experimenting With the Connections
When the PCB board of the LCD screen was disconnected from the PCB circuit board the display output was turned off.
When the white wire connecting the 16 tube PCR block to the PCB circuit board ability to regulate the temperature of the PCR was lost.
After finishing the diagnostic analysis, the PCR was tested by setting thermal cycler program to three stages. Stage one was one cycle of 95 °C for 3 minutes, the second stage was 35 cycles, 95 °C for 30 seconds, 50 °C for 30 seconds, 72 °C for 30 seconds, stage three was one cycle of 72 °C for 3 minutes. The test run lasted for about an hour and thirty minutes and confirmed that the temperature readings on the LED of the PCR machine and the computer matched.
Polymerase Chain Reaction
(Add your work from Week 3, Part 1 here)
(Add your work from Week 3, Part 2 here)
Research and Development
Specific Cancer Marker Detection - The Underlying Technology
The r17879961 sequence will produce a cancer mutation at Chromosomes 22 of the gene sequence. The normal sequence has a T ( thymine) nucleotide at chromosome 22 while the mutation sequence has an associated C (cytosine) nucleotide. The Open PCR machine is able to determine whether or not the r17879961 sample has cancer by replicating the desired mutation exponentially. Positive and negative strands are inserted into the PCR with a certain primer. The primer in the reaction is designed to attach to the C nucleotide that signifies cancer mutation. One strand has the primer, while the other strand does not. Open PCR will replicate the strand with the certain primer, causing an exponential growth. The negative strand will grow in a linear fashion. The PCR process goes through 30 cycles to complete this. After the PCR process, fluorescent dye is added to the solutions. The fluorescent dye will cause the DNA with double strands to glow. Since the PCR has grown the double stranded positive DNA exponentially the fluorescent dye glows brighter. Therefore the cancer DNA is in the sample with the glow.
(Your group will add the results of your Fluorimeter measurements from Week 4 here)