LAB 5 WRITE-UP
PCR Reaction Report
Our team was able to use the pipette successfully and had no trouble with it during part C of our lab. The pre-lab video did help us due to the fact that explained how to use it thoroughly and each one of us knew how to do so. The first stop on the pipette allows you to retrieve liquid or any substance you are trying to reach inside of the pipette. The second stop is when you are releasing the material in the pipette into a tube, cylinder, etc. Our team understood this from the pre-lab reading and had no trouble understanding it. The final reactions did have the same amount of liquid. After using the pipette to combine the DNA samples and PCR reaction mix, each tube we had labeled had the same amount of liquid. In addition, there was no liquid left in either of the tubes that contained DNA samples and PCR reaction mix. The labeling scheme we used was provided in the lab workbook; we used the same names the workbook had to label our samples so that we did not have to change our labeling scheme.
Smart Phone Camera Settings
- Type of Smartphone: Iphone 5S
- Flash: not on
- ISO setting: automatic
- White Balance: automatic
- Exposure: automatic
- Saturation: automatic
- Contrast: automatic
Place the camera in the cradle at a right angle to the slide. Make sure the camera views the droplet from the side and is at least 4 centimeters away from the droplet.
- Distance between the smart phone cradle and drop =
7 cm between the phone cradle and drop.
Placing Samples onto the Fluorimeter
- [Instructions: Step one, in your OWN words]
- [Instructions: Step two, in your own words]
- [Instructions: Step three, in your own words]
- [Instructions: Step etc., in your own words]
Data Collection and Analysis
Images of High, Low, and Zero Calf Thymus DNA
Calibrator Mean Values
|Initial Concentration of 2X Calf Thymus DNA solution (micrograms/mL)
||Final DNA concentration in SYBR Green I solution (µg/mL)
||RAWINTDEN DROP - BACKGROUND
|| Standard Deviation
Images of Our PCR Negative and Positive Controls
PCR Results: PCR concentrations solved
|PCR Product TUBE LABEL
||MEAN (of RAWINTDEN DROP - BACKGROUND)
||PCR Product Concentration (µg /mL)
||Initial PCR Product Concentration (µg /mL)
| G14 +
| G14 -
| G14 1-1
| G14 1-2
| G14 1-3
| G14 2-1
| G14 2-2
| G14 2-3
PCR Results: Summary
- Our positive control PCR result was 28.37 μg/mL
- Our negative control PCR result was 4.67 μg/mL
- Patient 69477 : The image is a circular object with light white shade but more dark unlike the first one. The observed quantitative data was 28.37 ug/ml.
- Patient 37835 : The image is a ciruclar object that is shaded very dark and has a quantitative number of 4.67 ug/ml.
- Patient 69477 : Negative. The initial values came out negative, so no PCR product was there.
- Patient 37835 : Negative. The initial values came out negative, so no PCR product was there.