BME100 f2013:W1200 Group13 L5: Difference between revisions
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'''How the Fluorescence Technique Works'''<br> | '''How the Fluorescence Technique Works'''<br> | ||
''The first step is to turn on the Fluorimeter, and place it on a flat surface such as a table or lab bench. Then place the glass slide with the smooth side down into the slot on top of the Fluorimeter, once this is completed adjust the height to get the camera placement correct where the slide view is nearly edge-on. After setting up the device, place the 80 uL drop of SYBR Green solution between the first two circles in the middle of the slide. After adding the 80uL of SYBR Green, add 80uL of DNA solution on top of the green dye. Once both solutions are on the slide adjust the slide so that the light passes through the center and to the other side. The distance between the camera and Fluorimeter should be greater than 4cm, if so you can then cover the device with black box keeping one flap up until camera is ready. Set the timer on camera and once it is focused lower the flap to block out the outside light before the picture is taken. Now the data is recorded and ready for analysis on ImageJ, so remove the 160uL drop from the slide and discard in proper waste container. '' | ''The first step is to turn on the Fluorimeter, and place it on a flat surface such as a table or lab bench. Then place the glass slide with the smooth side down into the slot on top of the Fluorimeter, once this is completed adjust the height to get the camera placement correct where the slide view is nearly edge-on. After setting up the device, place the 80 uL drop of SYBR Green solution between the first two circles in the middle of the slide. After adding the 80uL of SYBR Green, add 80uL of DNA solution on top of the green dye. Once both solutions are on the slide adjust the slide so that the light passes through the center and to the other side. The distance between the camera and Fluorimeter should be greater than 4cm, if so you can then cover the device with black box keeping one flap up until camera is ready. Set the timer on camera and once it is focused lower the flap to block out the outside light before the picture is taken. Now the data is recorded and ready for analysis on ImageJ, so remove the 160uL drop from the slide and discard in proper waste container. '' <br> | ||
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''The SYBR Green dye works as an indicator for the presence of DNA because it only displays fluorescence in the presence of double stranded DNA molecules. In the vials where PCR was successful, higher concentrations of DNA should result in more expression of the SYBR Green fluorescence.'' | |||
Revision as of 12:23, 13 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
ProcedureSmart Phone Camera Settings
Steps for Calibration
Solutions Used for Calibration [Instructions: See worksheet page 6.]
Placing Samples onto the Fluorimeter
Data AnalysisRepresentative Images of Samples DNA Positive Signal Picture Image J Values for All Samples
Fitting a Straight Line
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