BME100 f2016:Group8 W1030AM L5

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OUR TEAM

Name: Ryan Fuller
Name: Kasun Daundasekara
Name: Morgan Stephen
Name: Nick Keiper
Name: Brandon Davison


LAB 5 WRITE-UP

PCR Reaction Report

Yes the pre-lab reading helped our group understand what was going on in the lab. The first and second stop were easily figured out, and our group had no problems regarding measuring out the samples and distributing them. The final reactions did not have exactly the same amount of liquid in them due to a small amount of human error but they were very close. There was a very small amount of liquid left inside the tubes that it was inconsequential to the results of this lab. We did not change our labeling scheme throughout the experiment.

Fluorimeter Procedure

Imaging set-up

-First our group placed the sample onto the slide and turned on the laser. Next, we changed the settings on the Iphone used to match the settings in the lab workbook. Then, we measured out a good distance of approximately 12 cm from the fluorimeter and placed the Iphone on a stand there. The timer on the phone was then set for 5 seconds and the cover was quickly placed on the fluorimeter so the picture could be taken. Lastly, we repeated these steps 3 times for each drop that we placed on the slide.


Placing Samples onto the Fluorimeter

  1. [Step 1: Put a new tip onto the pipette.]
  2. [Step 2: Gather 80 micro liters of the DNA solution and place it on the flourimeter. ]
  3. [Step 3: Discard the tip used and place a new tip on the pipette. ]
  4. [Step 4: Gather 80 micro liters of the SYBR green solution and mix it with the DNA solution to form one drop. ]
  5. [Step 5: Repeat steps 1-4 for each new DNA solution.]


Data Collection and Analysis

Images of High, Low, and Zero Calf Thymus DNA

5 μg/mL sample 5 sample.jpg

0.5 μg/mL sample 0.5 sample.jpg

Zero DNA sample No DNA sample.jpg



Calibrator Mean Values


Table.png

Calibration curves

Plot 1.png

Plot 2.png

Images of Our PCR Negative and Positive Controls

Negative Negative Drop.jpg

Positive Positive image.jpg


PCR Results: PCR concentrations solved

Screen Shot 2016-11-08 at 10.01.54 PM.png


PCR Results: Summary

  • Our positive control PCR result was 82.95015829 μg/mL
  • Our negative control PCR result was _19.19627429_ μg/mL


Observed results

  • Patient 40211 : -16.04346686:
  • Patient 31028: 19.56305943:


Conclusions

  • Patient 40211 :Inconclusive. Our results were inaccurate due to some mistakes made during the procedure. We therefore can not confidently say whether the patient was positive or negative.
  • Patient 31028 :Negative. Although these results were also questionable, one of the results matches up to the negative control.