BME100 f2015:Group17 8amL5

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Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
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Contents

OUR TEAM

Name: Sam Stone
Name: Sam Stone
Name: Jadyn Ziegler
Name: Jadyn Ziegler
Name: Mahoro Uwiringiyimana
Name: Mahoro Uwiringiyimana
Name: Jake Feltz
Name: Jake Feltz
Name: Zachary Ramsey
Name: Zachary Ramsey
Name: Nathan Natividad
Name: Nathan Natividad


LAB 5 WRITE-UP

PCR Reaction Report

Our experience with micropipetting the samples in this reaction were effective. We were able to successfully extract accurate amounts of microliters onto the slidess in order to be tested. The pre-lab was useful because it was able to teach our group how to efficiently pipette accurate amounts to use in our lab. We were able to understand the difference between the first and second stops while also understanding their function for pipetting. The final reactions did have the same amount of liquid as the others and there was some remaining liquid in the tubs that held the samples but that is only because we used the correct amount of microliters in our measurements. We did not have to change our labeling scheme. All we did was simply lay the PCR samples on the table in the order that we wanted to test them in and we were able to organize the data that way.

Fluorimeter Procedure

Smart Phone Camera Settings

  • Type of Smartphone: HTC 1 M8
    • Flash:No
    • ISO setting:N/A
    • White Balance: N/A
    • Exposure:N/A
    • Saturation:N/A
    • Contrast:N/A


Camera set-up
A metal stand was utilized to hold the phone horizontally in place. The Phone slipped to an incorrect angle, so a spacer was created out of plastic to hold the phone at a ninety degree angle.

  • Distance between the smart phone cradle and drop =

Distance in centimeters: 8.0 centimeters

For concentration 5, distance was 9.5 cm

Concentration 2, distance was 11 cm

Concentration 1, distance was 11 cm

Concentration .5, distance was 11 cm

Once we tested the DNA samples, centimeter length was 11.5


Placing Samples onto the Fluorimeter

  1. Step one, set the micropipettor to 80 microliters
  2. Step two, extract 80 microliters of the SYBR Green and place onto the slide in the fluorimeter
  3. Step three, Get a new tip, then extract 80 microliters of the DNA and place on the same drop of the SYBR Green
  4. Step four, Place phone in a proper angle in front of the fluorimeter and take a picture with the box cover closed over it
  5. Step five, take three pictures of each DNA sample
  6. Step six, clean up the sample on the slide and dispose of the tip and sample, then move the slide in the fluorimeter to a new positions
  7. Step seven, repeat steps listed above for each of the different DNA samples


Data Collection and Analysis

Images of High, Low, and Zero Calf Thymus DNA High photo


Low photo


Zero photo



Calibrator Mean Values

Image:bme1001.png‎ TABLE GOES HERE


Calibration curves

Image:bme1011.png‎
 Description of image


Images of Our PCR Negative and Positive Controls Positive photo


Negative photo



PCR Results: PCR concentrations solved

secondchart


PCR Results: Summary

  • Our positive control PCR result was 96 μg/mL
  • Our negative control PCR result was 86 μg/mL


Observed results

  • Patient 97780: Because this patient was positive the SYBR Green portrayed a droplet that glowed green. The average quantitative description of this patient was about 82 (ug/mL)
  • Patient 21836:Because this patient was negative, the SYBR Green droplet was a clear and transparent drop. The average quantitative description of this patient was 92.66 (ug/mL)


Conclusions

  • Patient 97780: Although the image shows that it was positive due to the presence of the green color under fluorescence light, the numbers don't agree with it because the concentration was closer to the negative control than it was closer to the positive control.
  • Patient 21836: Like patient 97780, this patient's value did not agree with the observation during experiment. The color showed negative result, but the values were closer to the positive than they are to the negative. This could have been a result of the calculation error that resulted in the unexpected values. Therefore, one cannot conclude that the patient was positive or negative.



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