BME103 s2013:T900 Group6
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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
The Original Design
When we unplugged the LCD screen from the OpenPCR circuit board, the machine's LED failed to work properly.
When we unplugged the white wire that connects the OpenPCR circuit board to the main heating block, the machine began to show a change of the temperature on the LCD screen.
The date the PCR machine was open to conduct an analysis was on March 5th, 2013. We had machine number 6 that day. What we came to see, in terms of pros and cons, contained the following:
Pros: - Lightweight - Quiet - Ease of Use
- Took a while to complete the full cycle
- The lid is not easy to remove. Requires some work
- Requires a computer
- Small and somewhat heavy
- Made of Wood (fire hazard)
Thermal Cycler Program
DNA Sample Set-up
DNA Sample Set-up Procedure 1. Obtain DNA samples, PCR, DNA/Primer vials 2. Open All The Tops 3. Use pipette to move DNA into vials containing PCR mix
PCR Reaction Mix
The PCR mix is made up of 50 Microliters of PCR.
DNA/ primer mix
The DNA/Primer Mix is made of 50 micro liters of DNA of patients 1 and 2, and primer. There are 2 vials containing negative and positive control. The negative control does not have cancer DNA, and the positive control has cancer in it.
Research and Development
How PCR Replicates DNA
What is DNA? DNA, short for deoxyribonucleic acid, is located within the nucleas of a cell. DNA is shaped as a double helix with two strands on each side, some refer to it as a winding staircase. In between the strands there are things called nucleotides, or the stairs, which hold the DNA together. There are four different kinds of nucleotides that form base pairs when they are bonded together. These base pairs are Guanine-Adenine and Thymine-Cytosine.
The Process The PCR is needed in the replication of DNA based on the fact that the PCR machine has the ability to heat and cool down the substances that are placed inside of it. These temperature changes are crucial in the replication process because at high temperatures DNA strands seperate into single strands. Before the tube is even placed in the PCR machine you will need more then just DNA inside the tube. Primers is the first thing to interact with the DNA. While the DNA is separated into two strands, primers, which are pretty much short single strands of DNA, bind to the DNA instead of letting the DNA bind together to itself again. This is where the nucleotide base pairs come in, each primer has the exact nucleotide sequence for the DNA in order to bind to it. The primers are to short to finish the whole DNA replication, polymerase comes in to finish the job with the help of magnesium chloride and dNTP, which just help grab nucleotides for the polymerase. The replication process happens over and over again due to the temperature control of the PCR machine until you have enough DNA to be observed.
Once again, in our experiment, we used the PCR machine, also known as the polymerase chain reaction machine. It is used to amplify strands of DNA and generate many copies of a specific sequence. As previously stated, it can be used to research different diseases, especially cancer.