BME103 s2013:T900 Group1 L2: Difference between revisions
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'''Calibration'''<br> | '''Calibration'''<br> | ||
In order to measure unknown concentrations of DNA with the fluorimeter by measuring fluorescence, we must first measure the fluorescence of known concentrations of DNA using the same technique. To accomplish this, we used an assay of several known concentrations of calf Thymus DNA in conjunction with SYBR green dye. We took pictures of the fluorescing drops and used imaging software to measure the fluorescence. SYBR green dye fluoresces green and the | In order to measure unknown concentrations of DNA with the fluorimeter by measuring fluorescence, we must first measure the fluorescence of known concentrations of DNA using the same technique. To accomplish this, we used an assay of several known concentrations of calf Thymus DNA in conjunction with SYBR green dye. We took pictures of the fluorescing drops and used imaging software to measure the fluorescence. SYBR green dye fluoresces green and the ImageJ software allows the colors of one picture to be split into pictures of the component colors. We can then select just the green light and measure the pixel density, allowing us to quantify the fluorescence. With a known concentration and a known pixel density, we can then define a relationship between the two which will allow us to later measure the fluorescence of an unknown sample to determine the DNA concentration of it. <br> | ||
We inserted the iPhone inside a cradle that gave us a right angle view. Afterward we adjusted the height of the phone so that it is as even with the drop. | We inserted the iPhone inside a cradle that gave us a right angle view. Afterward we adjusted the height of the phone so that it is as even with the drop on the slide. | ||
[[Image:Photo(1).JPG]] | [[Image:Photo(1).JPG]] | ||
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'''Solutions Used for Calibration' | '''Solutions Used for Calibration''' | ||
{| {{table}} width=700 | {| {{table}} width=700 | ||
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'''Placing Samples onto the Fluorimeter''' | '''Placing Samples onto the Fluorimeter''' | ||
* '' | * ''Align light between first two clean rows of spots so it will shine directly at the drop, secure'' | ||
* '' | * ''Using a clean micro-pipette tip, slowly place 80μL of SYBR green dye allowing it to span between both spots'' | ||
* '' | * ''Using a clean micro-pipette tip, add 80μL of the sample solution to the SYBR dye'' | ||
* '' | * ''Complete assembly by placing the camera/cradle in position and cover with light box'' | ||
* ''Start timed delay on phone camera'' | |||
* ''Close light box quickly and carefully and wait the allotted time for the picture to complete'' | |||
* ''Extract and discard solution from the slide'' | |||
These steps were repeated in triplicate for each of the samples listed above. | |||
<br> | <br> |
Revision as of 21:00, 1 April 2013
BME 103 Spring 2013 | Home People Lab Write-Up 1 Lab Write-Up 2 Lab Write-Up 3 Course Logistics For Instructors Photos Wiki Editing Help | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
OUR TEAM
LAB 2 WRITE-UPBackground InformationSYBR Green Dye Single-Drop Fluorimeter
ProcedureSmart Phone Camera Settings
Calibration
These steps were repeated in triplicate for each of the samples listed above.
Data AnalysisRepresentative Images of Samples
Image J Values for All Samples [See worksheet page 5]
[Add more rows as needed]
Fitting a Straight Line
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