BME103:W930 Group9: Difference between revisions
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Procedure for amplifying a person’s DNA <br> | Procedure for amplifying a person’s DNA <br> | ||
1. | 1. Add .2ng’s of a person’s dna to a tube containing the proper primers, water, and the GoTaq master mix. Repeat this step for as many replicates as you need. <br> | ||
2. | 2. Create a program on the open PCR machine that will run the proper cycle for replicating DNA. <br> | ||
3. The proper program will include an initial temperature hold of 95 degrees Celsius for three minutes, followed by 35 cycles of 95 to 57 to 72 all for 30 seconds each, then a final hold for 3 minutes at 72 degrees. <br> | 3. The proper program will include an initial temperature hold of 95 degrees Celsius for three minutes, followed by 35 cycles of 95 to 57 to 72 all for 30 seconds each, then a final hold for 3 minutes at 72 degrees. <br> | ||
4. Place the tubes in the open pcr machine making sure to close the lid. <br> | 4. Place the tubes in the open pcr machine making sure to close the lid. <br> | ||
5. | 5. Run the program with the tubes inside the machine. <br> <br> | ||
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! Total Volume || 100.0 μL | ! Total Volume || 100.0 μL | ||
|}<br><br> | |}<br><br> | ||
The | '''The Samples'''<br> | ||
The eight samples run in our pcr experiment were a positive control, a negative control, and three replicate samples from each of our patients.<br> | The eight samples run in our pcr experiment were a positive control, a negative control, and three replicate samples from each of our patients.<br> | ||
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Fluorimeter setup <br><br> | Fluorimeter setup <br><br> | ||
Flourimeter procedure<br><br> | '''Flourimeter procedure'''<br><br> | ||
1. | 1. Place the flourimeter on the table and turn on the blue light.<br> | ||
2. | 2. Place provided glass slide on flourimeter track so that the first row of dots is even with the light.<br> | ||
3. | 3. Place phone cradle in front of the flourimeter with a smart phone facing perpendicular to the beam of light. (as seen in the picture). <br> | ||
4. | 4. Add two drops of green dye on the dots that are even with the light.<br> | ||
5. Place two drops of DNA sample on top of the green dye.<br> | 5. Place two drops of DNA sample on top of the green dye.<br> | ||
6. | 6. Cover the flourimeter and phone by turning the large box over and placing it above both of them.<br> | ||
7. Take a picture of the droplet with the camera.<br> | 7. Take a picture of the droplet with the camera.<br> | ||
8. | 8. Save the picture and send it to the imageJ operator.<br> | ||
Instructions for opening images in imageJ<br><br> | Instructions for opening images in imageJ<br><br> |
Revision as of 23:04, 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 The open PCR is a device that enables the splitting of DNA. It is run through many cycles of heating and cooling that enables the splitting and recombination with polomers. It connects to a computer program to run the cycles. The temperature change is controled by the heating lid and than the information is fed to the LED screen. The heating lid heats tubes that are held in the heat tube holder to the appropriate temperature and then cools them as needed in each cycle. Experimenting With the Connections When we unplugged part 3, the LCD, from part 6, the Open PCR Brains Board, the LCD on the machine stopped working and did not show anything on it. When we unplugged the white wire that connects part 6, the Open PCR Brains Board, to part 2, the heat tube, the machine no longer measures the temperature of the plate and sends it to the LCD.
First test run: 24 Oct. 2012
ProtocolsPolymerase Chain Reaction Procedure for amplifying a person’s DNA
The Samples
Flourimeter procedure 1. Place the flourimeter on the table and turn on the blue light. Instructions for opening images in imageJ
Research and DevelopmentSpecific Cancer Marker Detection - The Underlying Technology This report used the technique called DNA amplification. (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
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