BME100 f2013:W900 Group13 L5: Difference between revisions
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# Before the sample can be placed on the Fluorimeter, the fluorimeter must be set up. This is done by first by placing the glass down on the fluorimeter; the glass must be placed so that the rough side is faced up. The fluorimeter must be turned on. Adjust the glass slide so that the blue light shines in the center of two rows. The hieght of the fluorimeter may need to be adjusted so that the camera has a clear view of the light. | # Before the sample can be placed on the Fluorimeter, the fluorimeter must be set up. This is done by first by placing the glass down on the fluorimeter; the glass must be placed so that the rough side is faced up. The fluorimeter must be turned on. Adjust the glass slide so that the blue light shines in the center of two rows. The hieght of the fluorimeter may need to be adjusted so that the camera has a clear view of the light. | ||
#For the next steps, the micropipet will be needed. Begin by adjusting the volume on the micropipet to 80μL, and placing a new micropipet tip on the micropipet. | #For the next steps, the micropipet will be needed. Begin by adjusting the volume on the micropipet to 80μL, and placing a new micropipet tip on the micropipet. | ||
# | # Use the micropipet to gather 80μL of the SYBR Green I to be placed on the slide. The SYBR Green I will be placed on the glass slide for each sample. Be sure that the camera settings are all in place and take photograph of solely SYBR Green I, this is your blank. | ||
# | # Next, use the micropipet to gather 80μL of the first sample, 0.25, and place it into the 80μL SYBR Green I present on the slide. Be sure that the light from the fluorimeter is in line with the droplet. | ||
# Photograph the sample and SYBR Green I. | |||
#Remove sample from slide with micropipet and repeat steps for each sample. After all samples have been collected (0,0.25,0.5,1,2,5) repeated this process two more times for a total of three pictures per sample. | |||
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Revision as of 16:17, 12 November 2013
BME 100 Fall 2013 | 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-UPBackground InformationSYBR Green Dye
How the Fluorescence Technique Works The fluorescence technique works by combining a fluorescent dye to the DNA. In this lab, SYBR Green dye was used as the fluorescent dye. The droplets will remain in place due to the Teflon coated glass. A beam of blue light is emitted from the single-drop fluorimeter and is passed through the droplet. A fluorescent green light will be emitted from the droplet which will be measured by taking a picture on a smartphone and analyzing the picture file on ImageJ.
ProcedureSmart Phone Camera Settings
Calibration 1. Adjust the camera settings on the smart phone so that they match the settings above. 2. Place smart phone in the cradle. 3. Make sure that the distance from the smart phone and the cradle is 11 cm. Solutions Used for Calibration
Data AnalysisRepresentative Images of Samples [Instructions: Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with no DNA] [Instructions: Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with DNA (positive signal)]
Image J Values for All Samples
Fitting a Straight Line [Instructions: Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 9]
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