LAB 5 WRITE-UP
PCR Reaction Report
The pre-lab reading was helpful because it explained how to effectively use the micro pipette. The pipette was used to get varied amounts of solutions for the experiment and we did understand difference between the first and second stop on the pipettor. The final reactions had the total amount of liquids that were mixed which was 80 and 80 from both of the solutions. There was no liquid leftover in the tubes that the DNA samples and PCR reactions mix and we did not have to change any labeling scheme.
1. Place the solutions in clear view, middle of the fluorimeter, so the camera can take a picture of it.
2. Measure distance between the solution and the camera and make sure the distance does not vary.
3. Place a box over the fluorimeter so that the solution is in darkness so the light can pass through without any interference.
4. Place a timer on phone to take 3 pictures in an allotted time.
5. Repeat steps until 3 pictures have been taken for all desired solutions.
Placing Samples onto the Fluorimeter
- 1. Take the pipette and take 80 microliters of the Green SYBR sample and place it in the middle of the fluorimeter in order to have the light pass through the drop.
- 2. Take the pipette and take 80 microliters of the calf thymus sample and place it on top of the previous solution.
- 3. Make sure the drop is aligned with the light and then place the box over the fluorimeter so no light passes through the light.
- 4. Grab phone and place a timer on it so it take 3 pictures of the drop as the light passes through it.
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 15.703 μg/mL
- Our negative control PCR result was 32.309 μg/mL
- Patient 45117 : The positive patient's solution had glowed and the concentration turned out to be 15.703 μg/mL
- Patient 82337 : The negative patient's solution did not glow and the concentration turned out to be 32.309 μg/mL
- Patient 45117: This patient is similar to the negative because the mean concentration of the negative is more similar to this patient.
- Patient 82337 :This patient is similar to the positive because the mean concentration of the positive is more similar to this patient.