BME103:T130 Group 2: Difference between revisions
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|+'''Polymerase Chain Reaction''' | |+'''Polymerase Chain Reaction'''<br> | ||
! '''Reagent'''!! '''Volume''' | ! '''Reagent'''!! '''Volume''' | ||
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<br> | <br> | ||
'''Sample Descriptions''' | '''Sample Descriptions''' | ||
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8. Press "Start" to run the PCR machine. <br> | 8. Press "Start" to run the PCR machine. <br> | ||
9. Record data.<br> | 9. Record data.<br> | ||
<br> | <br> | ||
'''Using a Fluorimeter'''<br> | '''Using a Fluorimeter'''<br> | ||
[[Image:PCR101.png]]<br> | [[Image:PCR101.png]]<br><br> | ||
1. Smart Phone Stand<br> | 1. Smart Phone Stand<br> | ||
2. Smart Phone<br> | 2. Smart Phone<br> | ||
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<br><br> | <br><br> | ||
'''Fluorimeter Assembly Procedures:'''<br> | '''Fluorimeter Assembly Procedures:''' | ||
<br> | |||
1. Adjust the settings on the smart phone: no flash, contrast on the lowest setting, saturation on the highest setting and exposure on the highest setting<br> | 1. Adjust the settings on the smart phone: no flash, contrast on the lowest setting, saturation on the highest setting and exposure on the highest setting<br> | ||
2. Place the slide (glass facing down) onto the device.<br> | 2. Place the slide (glass facing down) onto the device.<br> | ||
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8. Take picture. For best results, set a timer on the phone to be able to close the box and maximize darkness.<br> | 8. Take picture. For best results, set a timer on the phone to be able to close the box and maximize darkness.<br> | ||
9. Using the pipette marked for waste, remove the drops from the slide.<br> | 9. Using the pipette marked for waste, remove the drops from the slide.<br> | ||
10. Repeat steps 3 through 8 for all samples, each time moving down the slide. You can conduct five tests per slide; in other words you will need two slides total.<br> | 10. Repeat steps 3 through 8 for all samples, each time moving down the slide. You can conduct five tests per slide; in other words you will need two slides total.<br><br> | ||
'''ImageJ Set Up Procedures:'''<br> | '''ImageJ Set Up Procedures:''' | ||
<br> | |||
1. Download ImageJ onto a laptop using the ImageJ disk.<br> | 1. Download ImageJ onto a laptop using the ImageJ disk.<br> | ||
2. Save desired photos to desk top.<br> | 2. Save desired photos to desk top.<br> | ||
3. | 3. Open ImageJ <br> | ||
4. On the menu, select ANALYZE, and then select SET MEASUREMENTS. Now choose Area, Integrated Density, and Mean Grey Value.<br> | |||
5. Now you can upload photo to ImageJ<br> | |||
6. In order to analyze the pictures, you need to make the menu selection IMAGE. Now select COLOR. Finally, choose SPLIT CHANNELS. This will bring up three windows labeled blue, green, and red. Since SYBR green dye is green, you will want to analyze the photos under the window marked Green. Close the windows marked Red and Blue.<br> | |||
7. Now, using the oval tool, draw an oval around the drop and select ANALYZE and then select MEASURE. Write down the data given by ImageJ. <br> | |||
8. Using your laptops arrow keys, move the oval to a dark area in the picture to get data for the "noise." Again, select ANALYZE and then select MEASURE. Write down the data given by ImageJ.<br> | |||
9. Repeat steps 5 through 8 for all of the images. <br> | |||
4. Ready to work ImageJ<br> | 4. Ready to work ImageJ<br> | ||
Revision as of 20:07, 14 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 Polymerase Chain Reaction (PCR) machine is used to test for variations in nucleotides. It is simple, easy to use, and portable. For our purposes we used the PCR machine to test for cancer in patients. It was able to test up to sixteen test tubes at a time and took slightly more than an hour and a half to complete the cycles. It heats up in order to separate the DNA strand and then cools to allow primers to attach and new replicated strands to form. It then heats back up to repeat the same cycles.
Experimenting With the Connections When we unplugged the LCD screen from the PCR machine, the machine's display screen stopped working, but information continued to transmit to the computer. When we unplugged the white wire that connects the Thermocouple to the 16-tube PCR block, the machine's temperature measurement was disabled, in other words, it no longer kept track of heat readings.
On October 18th and 20th the PCR machine responded normally with no signs of malfunction. However, it took a little bit longer to complete the cycle; specifically, it took an hour and forty minutes in total. The machine LED display matched the computer screen which was a sign of success and proper function.
Protocols
Sample 1: Positive Control (contains cancer DNA template), Tube label: P
Using a Fluorimeter
Fluorimeter Assembly Procedures:
ImageJ Set Up Procedures:
Research and DevelopmentSpecific Cancer Marker Detection - The Underlying Technology
Some basic components involved in a PCR reaction are: r17879961
Results
Cancerous Drop
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