Name: Sultan Alanazi
Name: Tyler Mitchell
Name: Logan Martin
Name: Maryam Alsuwailem
Name: Amber Sogge
Name: Austin James
LAB 5 WRITE-UP
PCR Reaction Report
The PCR reaction went as according to plan outlined in the previous lab. The pipetting technique was performed correctly. All of the tubes were labeled as specified and the pipet tips were disposed after every use. The pre-lab reading helped us because it was the only reference we had as a team. The difference between the first and second step of the micropipettor was fully understood after actual use of the pipettor. No noticeable errors were made.
Smart Phone Camera Settings
- Type of Smartphone: Windows Phone
- ISO setting:1600
- White Balance:auto
- Distance between the smart phone cradle and drop = 8 cm
Placing Samples onto the Fluorimeter
- Begin by wearing proper lab attire. Proper lab attire includes a lab coat, splash proof lab goggles, and nitrile gloves.
- Turn on the fluorimeter.
- Insert a slide into the fluorimeter.
- Set up a phone to be on camera mode and place the camera on the cradle 8 cm (or what works best) from the fluorimeter. Record the position for reference.
- Turn on the blue light to the fluorimeter.
- With a micropipette, place 80 microliters of SYBR Green I on the slide.
- Pipette 80 microliters of the calf thymus sample solution on top of the SYBR Green I.
- Make sure the blue light from the fluorimeter is shining through the sample solution mixture.
- Carefully, take 3 pictures of the mixture on the slide.
- Repeat the same process for the other solutions.
Data Collection and Analysis
Images of High, Low, and Zero Calf Thymus DNA
High Calf Thymus DNA:
Low Calf Thymus DNA:
Zero Calf Thymus DNA:
Calibrator Mean Values
|Initial Concentration of 2x calf thymus DNA solution (micro-grams/mL)
||DNA Concentration in SYBR Green I Solution (ug/mL)
||Rewritten Drop-background Image 1
||Rewritten Drop-background Image 2
||Rewritten Drop-background Image 3
Images of Our PCR Negative and Positive Controls
PCR Results: PCR concentrations solved
|PCR Product TUBE LABEL
||RAWINTDEN DROP - BACKGROUND (Image 1)
||RAWINTDEN DROP - BACKGROUND (Image 2)
||RAWINTDEN DROP - BACKGROUND (Image 3)
|MEAN (of RAWINTDEN DROP - BACKGROUND)
||"PCR Product Concentration (µg /mL)(Step 5 calculation)
||Initial PCR Product Concentration(µg /mL)(Step 6 calculation)
PCR Results: Summary
- Our positive control PCR result was 24.06 μg/mL
- Our negative control PCR result was 2.334 μg/mL
- Patient 16274 : The results for patient 1 appeared to be positive. There was visible green in the drop. The values for the first patient were 22.24118455μg/mL, 29.66080598μg/mL, and 23.99324013μg/mL. The average value was 25.29841022μg/mL. The results are close to the positive control of 24.06 μg/mL.
- Patient 42480 : The results for patient 2 also appeared to be negative. There was no visible green in the drop. The values for the second patient were 12.62686058μg/mL, 3.919564507μg/mL, and 1.520242931μg/mL. The average value was 6.022222673μg/mL. Although one reading was a little higher than normal, the overall average is closer to the negative control of 2.334 μg/mL.
- Patient 16274 :This patient's results turned out positive after compared with the positive and negative control. The average value for patient 16274 was close to the positive control. Through observational analysis, the drop was green thus further confirming a positive reading.
- Patient 42480 :The results for this patient were negative as well. The average value for patient 16274 was close to the negative control. Through observational analysis, the drop was not green thus further confirming a negative reading.