BME103 s2013:T900 Group5 L2

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(Procedure)
(Procedure)
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'''Calibration'''<br>
'''Calibration'''<br>
 +
The Camera was set up such that we could get the clearest image with maximum LED light absorption. A light box was placed over the fluorimeter to <br>
 +
darken the area. The Smart phone was aligned with the slide to get a sideways picture of the DNA sample. This creates the ideal angle to observe any <br>
 +
fluorescent light after the picture is taken. The camera phone is calibrated to view the fluorescent light with little interference from contrast and <br>
 +
recording the most light- done by setting the exposure high. The saturation is set to low so that color is not rendered in the photo. the ISO is set to <br>
 +
high for maximum focus in a dark environment.
-
''[Describe how to set up your camera in front of the fluorimeter. Add a PHOTO of this set-up for bonus points.]''
 
* Distance between the smart phone cradle and drop =
* Distance between the smart phone cradle and drop =
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'''Solutions Used for Calibration'''  
'''Solutions Used for Calibration'''  
{| {{table}} width=700
{| {{table}} width=700
-
| "Concentration Calf Thymus" <br> "(μg/mL)" || "Volume DNA added" <br> "(μL)"|| "Volume SABR GREEN I" <br> "(μL)" || "Concentration resulting DNA"<br> "(ng/mL)"
+
| Concentration Calf Thymus <br> (μg/mL) || Volume DNA added <br> (μL)|| Volume SABR GREEN I <br> (μL) || Concentration resulting DNA<br> (ng/mL)
|-
|-
| 5 || 80 || 80 || 2.5
| 5 || 80 || 80 || 2.5
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'''Placing Samples onto the Fluorimeter'''
'''Placing Samples onto the Fluorimeter'''
-
* Step 1.) With the Camera set in place we prepared the slide with SYBR Green I, a constant throughout the experiment <br>
+
* 1.) With the Camera set in place we prepared the slide with SYBR Green I, a constant throughout the experiment <br>
-
* Step 2.) 80 μL of 5μg/mL of calf thymus concentration DNA samples were then added to the slide <br>
+
* 2.) 80 μL of 5μg/mL of calf thymus concentration DNA samples were then added to the slide <br>
-
* Step 3.) To take 3 pictures in the the dark with as much of the blue LED light as possible, we used a timer on the camera, set to 3 seconds. <br>
+
* 3.) To take 3 pictures in the the dark with as much of the blue LED light as possible, we used a timer on the camera, set to 3 seconds. <br>
-
* Step 4.) The slide, now containing SYBER Green I and Calf Thymus DNA was then aligned in the cradle such that the light was shining into the DNA. <br>
+
* 4.) The slide, now containing SYBER Green I and Calf Thymus DNA was then aligned in the cradle such that the light was shining into the DNA. <br>
-
* Step 5.) With the Camera phone, we captured 3 images of the DNA and removed the light box. <br>
+
* 5.) With the Camera phone, we captured 3 images of the DNA and removed the light box. <br>
-
* Step 6.) That sample was then disposed of with a micropipette <br>
+
* 6.) That sample was then disposed of with a micropipette <br>
-
* Step 7.) Repeat with 80μL Green SABR I and 80μL of each Calf Thymus Concentration given by the first column in the table above<br>
+
* 7.) Repeat with 80μL Green SABR I and 80μL of each Calf Thymus Concentration given by the first column in the table above<br>
-
<!-- Note: Be sure to delete the text in brackets: ''[ ]'' -->
+
==Data Analysis==
==Data Analysis==

Revision as of 01:53, 2 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
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Image:BME494_Asu_logo.png


Contents

OUR TEAM

Name: Alexander Oropel
Name: Alexander Oropel
Name: Matt McClintock
Name: Matt McClintock
Name: student
Name: student
Name: student
Name: student
Name: student
Name: student
Name: student
Name: student


LAB 2 WRITE-UP

Background Information

SYBR Green Dye
[A short summary describing SYBR green dye]


Single-Drop Fluorimeter
[A description of the single-drop fluorimeter device. Add a PHOTO for bonus points]


How the Fluorescence Technique Works
[In your own words, a summary of the information from page 9 of the worksheet]



Procedure

Smart Phone Camera Settings

  • [The type of smart phone you used and how you adjusted the camera settings, if applicable (see worksheet page 4).]
    • Flash:
    • ISO setting:
    • White Balance:
    • Exposure:
    • Saturation:
    • Contrast:
  • [If you used an additional phone, describe the other type of smart phone you used and how you adjusted the camera settings, if applicable (see worksheet page 4).]
    • Flash:
    • ISO setting:
    • White Balance:
    • Exposure:
    • Saturation:
    • Contrast:


Calibration
The Camera was set up such that we could get the clearest image with maximum LED light absorption. A light box was placed over the fluorimeter to
darken the area. The Smart phone was aligned with the slide to get a sideways picture of the DNA sample. This creates the ideal angle to observe any
fluorescent light after the picture is taken. The camera phone is calibrated to view the fluorescent light with little interference from contrast and
recording the most light- done by setting the exposure high. The saturation is set to low so that color is not rendered in the photo. the ISO is set to
high for maximum focus in a dark environment.


  • Distance between the smart phone cradle and drop =

[See worksheet page 5]


Solutions Used for Calibration

Concentration Calf Thymus
(μg/mL)
Volume DNA added
(μL)
Volume SABR GREEN I
(μL)
Concentration resulting DNA
(ng/mL)
5 80 80 2.5
2 80 80 1
1 80 80 0.5
0.5 80 80 0.25
0.25 80 80 .125
0 80 80 blank


Placing Samples onto the Fluorimeter

  • 1.) With the Camera set in place we prepared the slide with SYBR Green I, a constant throughout the experiment
  • 2.) 80 μL of 5μg/mL of calf thymus concentration DNA samples were then added to the slide
  • 3.) To take 3 pictures in the the dark with as much of the blue LED light as possible, we used a timer on the camera, set to 3 seconds.
  • 4.) The slide, now containing SYBER Green I and Calf Thymus DNA was then aligned in the cradle such that the light was shining into the DNA.
  • 5.) With the Camera phone, we captured 3 images of the DNA and removed the light box.
  • 6.) That sample was then disposed of with a micropipette
  • 7.) Repeat with 80μL Green SABR I and 80μL of each Calf Thymus Concentration given by the first column in the table above

Data Analysis

Representative Images of Samples

[Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with no DNA]

[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 [See worksheet page 5]

[Add more rows as needed]Fitting a Straight Line
[Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 8]
row 1 cell 1 row 1 cell 2 row 1 cell 3 row 1 cell 4 row 1 cell 5
row 2 cell 1 row 2 cell 2 row 2 cell 3 row 2 cell 4 row 2 cell 5
row 3 cell 1 row 3 cell 2 row 3 cell 3 row 3 cell 4 row 2 cell 5
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