OUR TEAM
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
- Make sure that the fluorimeter is on, with the camera in the correct spot with the correct settings.
- Place an 80 mL drop of the SBYR Green sample in between the first two dots on the slide.
- Place an 80 mL drop of the desired sample on the preexisting drop. Ensure that the light penetrates the center of the drop.
- 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
Error creating thumbnail: File missing
Error creating thumbnail: File missing
Error creating thumbnail: File missing
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
|