LAB 5 WRITE-UP
PCR Reaction Report
Pipetting was a relatively simple task to get accumulated to. Initially, we were relatively inefficient when it came down to pipetting, but by the end of the lab, we were adding samples, and taking pictures at a much faster rate while still being accurate. The pre-lab reading was very effective when it came down to helping us use the pipettor. The difference from the first and second stop was very easy to figure out, and utilize when taking and placing samples. The final reactions did have the exact same amount of liquid, due to the accuracy of the pipettor, and we were able to use up all of the DNA samples. We did not have to modify our labeling scheme.
Smart Phone Camera Settings
- Type of Smartphone: iPhone 6
- ISO setting: 9
- White Balance: normal
- Exposure: normal
- Saturation: none added
- Contrast: none added
The camera was set up by placing it in the cradle provided in the materials, as well as placing the cradle on a additional surface to raise the camera up in order to take a better picture. After being placed in the cradle the phone is pointed at the Fluorimeter and set up to take pictures of the droplet.
- Distance between the smart phone cradle and drop = 5-6 cm
Placing Samples onto the Fluorimeter
- Slide a glass slate into Fluorimeter so that the smooth side is facing down
- Place 80 microliters of the SYBR Green 1 solution between the first 2 circles in the middle of the slide
- Add 80 microliters of the sample on the SYBR Green 1 drop
- Move the slate so that the light being emitted from the fluorometer is passing through the solution
Data Collection and Analysis
Images of High, Low, and Zero Calf Thymus DNA
Calibrator Mean Values
Images of Our PCR Negative and Positive Controls
PCR Results: PCR concentrations solved
PCR Results: Summary
- Our positive control PCR result was -19.21 μg/mL
- Our negative control PCR result was -20.52 μg/mL
- Patient 22993 : Quantitatively, on average values were closer to the negative control value. Qualitatively, they had the same appearance as the negative control, containing very similar characteristics such as being very dark with the periodic spot of brightness.
- Patient 85311 : Quantitatively, the average values were closer to the negative control value. Qualitatively, they had the same appearance as the negative control, containing very similar characteristics such as being very dark with the periodic spot of brightness
- Patient 22993 : For the second and third trials, the PCR concentrations were closer to the negative control. For the first trial, the value was closer to the positive control. Since the majority of values were closer to the negative, patient 22993 was negative
- Patient 85311 : For the first two trials, the PCR concentration values were closer to the negative control. For the third trial, the value was closer to the positive control. Since the majority of the trials were closer to the negative control, patient 85311 was negative.