BME103:T130 Group 9: Difference between revisions
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Eight samples were tested during this investigation. These included: a positive control with the cancer DNA template, a negative control without the cancer DNA template, and three samples each from the two subjects. The first subject was a 46 year old woman, correlating to test tubes labeled 2, and the other was a 62 year old man, correlating to test tubes labeled 4.<br> | Eight samples were tested during this investigation. These included: a positive control with the cancer DNA template, a negative control without the cancer DNA template, and three samples each from the two subjects. The first subject was a 46 year old woman, correlating to test tubes labeled 2, and the other was a 62 year old man, correlating to test tubes labeled 4.<br> | ||
''' | '''Fluorimeter Setup and Measurements'''<br> | ||
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| [[Image:FluorimeterGroup9Pic1.jpg|thumb|The basic components of the fluorimeter.]]<br> | | [[Image:FluorimeterGroup9Pic1.jpg|thumb|The basic components of the fluorimeter.]]<br> | ||
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In order to set up the fluorimeter, the following steps must be taken:<br><br> | |||
1. Remove the all the contents from the box. Open one of the small sides like a flap and place the box upside down on surface without the lid on. This will make the picture dark so only the blue light of the fluorimeter appears.<br> | |||
2. Place the superhydrophic slide in the fluorimeter, making sure the glass side is down and to align a row of holes with the light source.<br> | |||
3. Using a water dropper, place two to four droplets of water gently over the middle hole of the aligned row. <br> | |||
4. Place two to four droplets of the PCR solution for analysis into this water droplet. Be sure to be gentle as to not break surface tension. <br> | |||
5. Turn the light source on and place the fluorimeter in the box near the end of it that is already set up. <br> | |||
6. Place a smartphone with a good camera into the camera holder closer to the entrance of the box-cave.<br> | |||
7. Place your finger where the button to take a photo is and close the flap to reduce the amount of light from the outside coming in. Take a picture of the blue light shining through the solution. <br> | |||
8. Take several photos per solution being tested in order to get more stable data per that one solution of DNA being tested. <br> | |||
8. Repeat steps 2 through 8 for each solution of DNA used. <br><br> | |||
The next step is to analyze the resulting pictures by measuring the amount of pixels created by the light and comparing it to the controls. In order to measure this, the pictures must be analyzed using ImageJ software: <br> | |||
1. Download the ImageJ software from [http://rsbweb.nih.gov/ij/download.html] <br> | |||
2. Email the photos to yourself and download them onto the computer with ImageJ. <br> | |||
3. Open up ImageJ and go to file > import > image sequence. Determine the numbers of the images in your folder based upon their position alphabetically and place this in the starting number. Make sure to only have 1 in the number of images. <br> | |||
4. Repeat step 3 for every picture taken, using the differing numbers associated with each image to import them. <br> | |||
==Research and Development== | ==Research and Development== |
Revision as of 23:04, 7 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 the LCD from the Circut Board, the display was disconnected and would not appear.The PCR would still work. When we unplugged the white wire that connects the PCB Circut Board to Temperature Sensor, the machine would not be able to sense temperature correctly and would not give us a reading. Test Run (Write the date you first tested Open PCR and your experience(s) with the machine)
ProtocolsPolymerase Chain Reaction
Eight samples were tested during this investigation. These included: a positive control with the cancer DNA template, a negative control without the cancer DNA template, and three samples each from the two subjects. The first subject was a 46 year old woman, correlating to test tubes labeled 2, and the other was a 62 year old man, correlating to test tubes labeled 4. Fluorimeter Setup and Measurements
The next step is to analyze the resulting pictures by measuring the amount of pixels created by the light and comparing it to the controls. In order to measure this, the pictures must be analyzed using ImageJ software: Research and DevelopmentSpecific Cancer Marker Detection - The Underlying Technology Polymerase Chain Reaction (PCR) is a process by which we replicate DNA in order to determine specific sequences of DNA. In our specific lab, we used PCR to look for a sequence of nucleotides that signify cancer. On the surface level, we start with 2 strands of DNA, Magnesium Chloride (MgCl2), a TAQ enzyme, and DNTP, which is a mixture of the four bases – A, C, T, and G. We then set the PCR test to change temperatures after set periods of time to allow specific processes to work on the molecular level. We also add a fluorescent die that will bind to only the double strand of DNA. (BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the OpenPCR tutorial might be useful. Be sure to credit the source if you borrow images.)
Results(Your group will add the results of your Fluorimeter measurements from Week 4 here)
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