BME100 f2015:Group2 8amL5

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Owwnotebook icon.png BME 100 Fall 2015 Home
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Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
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OUR TEAM

Name: Bryce Goldthwaite
Role(s)
Name: Stephen Lane
Role(s)
Name: Samuel Rios
Role(s)
Name: Christian Manriquez
Role(s)
Name: Yash Dalvi
Role(s)


LAB 5 WRITE-UP

PCR Reaction Report

The Polymerase Chain Reaction went very well. The micropipetter was easy to use and understand. We began with labeling the tubes, which helped tremendously during the procedure. When there was any confusion, it was shortly resolved by slowing down for a second. There were a few tubes with a small amount of liquid left over and most of the tubes with mixes in them seemed to have about equal amounts in them.

Fluorimeter Procedure

Smart Phone Camera Settings

  • Type of Smartphone: iPhone 6
    • Flash: None
    • ISO setting: 2000
    • White Balance: Not Shown
    • Exposure: 2.2
    • Saturation: N/A
    • Contrast: N/A


Camera set-up
The iPhone 6 was placed in the camera cradle with the base of the cradle 9cm away from the fluorimeter. The cell phone was tilted forward in the cradle. The height of the fluorimeter was adjusted in order for the drop to be directly in front of the camera.

  • The distance between the base of the cradle and the base of the fluorimeter was 9cm.


Placing Samples onto the Fluorimeter

  1. Make sure that the fluorimeter is on, with the camera in the correct spot with the correct settings.
  2. Place an 80 mL drop of the SBYR Green sample in between the first two dots on the slide.
  3. Place an 80 mL drop of the desired sample on the preexisting drop. Ensure that the light penetrates the center of the drop.
  4. Cover the fluorimeter with the box, ensure that the camera settings are correct, lower the flap while pictures are taken.


Data Collection and Analysis

Images of High, Low, and Zero Calf Thymus DNA

  • 5 μg/mL

Also not water test.jpeg

  • 0.5 μg/mL

Water Test Group 2.jpg

  • Zero DNA

Not Water Test Group 2.jpeg

Calibrator Mean Values


Image number Final DNA concentration in SYBR Green I solution (µg/mL) AREA Mean Pixel Value RAWINTDEN OF THE DROP RAW INTDEN OF THE BACKGROUND
1 2.5 36828 35.213 1296816 109341
2 2.5 36828 37.519 1381768 134984
3 2.5 36828 36.907 1359195 139208
1 1 34852 74.005 2579210 122999
2 1 34852 74.898 2610348 112730
3 1 34852 74.542 2528242 120290
1 0.5 36640 114.73 4203711 151637
2 0.5 36640 115.034 4214841 159676
3 0.5 36640 114.548 4193752 161467
1 0.25 30584 122.628 3750467 131181
2 0.25 30584 122.645 3750970 33019
3 0.25 30584 122.209 3737636 123067
1 0.125 34254 173.018 5926549 126466
2 0.125 34254 173.119 5930012 122025
3 0.125 34254 173.046 4961784 128970
1 0 33975 179.438 6096593 163988
2 0 33975 180.133 6120200 166582
3 0 33975 179.131 6686168 162083

Calibration curves


Images of Our PCR Negative and Positive Controls


PCR Results: PCR concentrations solved

TABLE GOES HERE


PCR Results: Summary

  • Our positive control PCR result was 10.06 μg/mL
  • Our negative control PCR result was 1.443 μg/mL


Observed results

  • Patient 65991 : Bright green drops
  • Patient 29837 : Clear/blue drops


Conclusions

  • Patient 65991 : Positive
  • Patient 29837 : Negative