BME100 s2015:Group2 12pmL5: Difference between revisions
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'''Calibration'''<br> | '''Calibration'''<br> | ||
<!-- INSTRUCTIONS: In the space below, briefly describe how to set up your camera in front of the fluorimeter. Add a PHOTO of this set-up for bonus points. --> | <!-- INSTRUCTIONS: In the space below, briefly describe how to set up your camera in front of the fluorimeter. Add a PHOTO of this set-up for bonus points. --> | ||
* Camera should be set in the cradle at least 4 cm away from the drop placed on the slide | |||
* Camera should be set on a timer of three seconds in order to allow for a proper time of closing the box and no outside light to contaminate the pictures | |||
* The lens should be level with the drop (all of the drop's spherical surface from top to bottom should be in the picture) | |||
* Adjust Fluorimeter height if necessary to get it in the proper view of the phone camera | |||
<!-- INSTRUCTIONS: Type the distance between your phone cradle and the drop after the equal sign. --> | <!-- INSTRUCTIONS: Type the distance between your phone cradle and the drop after the equal sign. --> | ||
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[[Image:Bonus Question.png|400px|Bonus Question Picture]] | [[Image:Bonus Question.png|400px|Bonus Question Picture]] | ||
For the set-up, this picture represents what the set up looked like right before the group closed the flap to take the picture. The camera was set in the cradle and the lens was 6.8 cm away from the drop that was on the superhydrophic side of the glass slide. The box covered the | For the set-up, this picture represents what the set up looked like right before the group closed the flap to take the picture. The camera was set in the cradle and the lens was 6.8 cm away from the drop that was on the superhydrophic side of the glass slide. The box covered the Fluorimeter to protect the SyberGreen Dye from exposure to light. After close the flap, three pictures were taken using the timer on the iPhone to delay the taking of the pictures until after the flap was closed. | ||
'''Placing Samples onto the Fluorimeter''' | '''Placing Samples onto the Fluorimeter''' | ||
# | # Ensure that superhydrophic side of the slide is facing up when the slide is placed in the Fluorimeter | ||
# | # Turn On Fluorimeter by flipping the switch on the side of the Fluorimeter | ||
# | # Pipette 80 μL of SYBR Green I dye in between the first two clear circles on the glass slide (light from Fluorimeter should be absorbed by drop if the drop is in the proper place) | ||
# | # Replace the pipette tip and then pipette 80 μL of the PCR sample mix or the calibration sample on top of the SYBR Green I dye droplet | ||
# Make sure the light shines through the center of the drop and the light is focused through the opposite side of the drop | |||
# Ensure the iPhone rests in the cradle at least 4 cm away from the droplet | |||
# Set phone timer for 3 seconds and adjust camera as described in above calibration section | |||
# Make sure the iPhone camera focuses on the droplet | |||
# Cover the Fluorimeter with the box and press the button to take the picture on the iPhone and quickly close the flap to block all outside sources of light | |||
# Take at least three pictures of each sample tested to ensure that a good picture is obtained | |||
# Remove the used slide and dispose of properly | |||
# Repeat the above setup for each sample | |||
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Revision as of 16:03, 1 April 2015
BME 100 Spring 2015 | Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help | ||||||
OUR TEAM
LAB 5 WRITE-UPProcedureSmart Phone Camera Settings
For the set-up, this picture represents what the set up looked like right before the group closed the flap to take the picture. The camera was set in the cradle and the lens was 6.8 cm away from the drop that was on the superhydrophic side of the glass slide. The box covered the Fluorimeter to protect the SyberGreen Dye from exposure to light. After close the flap, three pictures were taken using the timer on the iPhone to delay the taking of the pictures until after the flap was closed. Placing Samples onto the Fluorimeter
Data AnalysisRepresentative Images of Negative and Positive Samples Negative Sample Positive Sample
Image J Values for All Calibrator Samples In the calibration section of the chart listed above, picture 1 correlates to H2O, picture 2 correlates to 0.25 units of DNA per microliter, picture 3 correlates to 0.5 units of DNA per microliter, picture 4 correlates to 1.0 units of DNA per microliter, picture 5 correlates to 2.0 units of DNA per microliter, and picture 6 correlates to 5.0 units of DNA per microliter. Picture 6 served as the positive control while picture 1 served as the negative control when checking the results of the PCR experiment.
Observed results
Conclusions
SNP Information & Primer DesignBackground: About the Disease SNP
Primer Design and Testing Diseased Primer <a href="http://imgur.com/ruzkEYS"><img src="http://i.imgur.com/ruzkEYS.png" title="source: imgur.com" /></a> NON-Diseased Primer <a href="http://imgur.com/TAV9kPT"><img src="http://i.imgur.com/TAV9kPT.png" title="source: imgur.com" /></a> The diseased primer had no matches because the humane genome does not have the mutation of AGT. Therefore, the would be no match with the natural human genome tested against in the website. This variation is what makes each individual's makeup unique. The AGT mutation is what makes a person have coronary heart disease.
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