BME100 f2016:Group14 W8AM L5

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Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
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OUR TEAM

Name: Diego Barra
Name: Savanna Bracale
Name: Casin Corallo
Name: Imelda Fragoza
Name: Andrew Samanta


LAB 5 WRITE-UP

PCR Reaction Report

The team's experience went very well with pipetting. Everyone in the team got to pipette and familiarize themselves by the end of the period. Yes, the pre-lab reading helped. The first stop is to pick up the solution and the second stop is to expel it. Yes, the final reactions had the same amount of liquid and there was not any liquid left in the tubes. We kept our labeling scheme the same.

Fluorimeter Procedure

Imaging set-up

To capture images from the flourimeter first the flourimeter was set up by turning it on andplacing a slide in with the smooth side down. We then, set up a phone timer to 3 seconds and turned off the flash. The phone went into a craddle and the flourimeter was lifted with boxes until the slide was in horizontal view of the camera.


Placing Samples onto the Fluorimeter

  1. Set micropippete to 80 uL
  2. Place tip on micropipette
  3. Press down on micropipette to first stop
  4. Place tip to SYBR Green I solution and release to pick up liquid
  5. Place tip between first 2 circles in the middle of the glass slide
  6. Press down to second stop on micropipette to release liquid onto slide
  7. Dispose of tip
  8. Place new tip on micropipette
  9. Press down on miropipette to first stop
  10. Place tip to sample/calibration solution and release to pick up liquid
  11. Place tip onto drop of SYBR Green I drop
  12. Press down to second stop to release the sample/calibration drop on top of the SYBR Green I drop



Data Collection and Analysis

Images of High, Low, and Zero Calf Thymus DNA

5 μg/mL sample
0.5 μg/mL sample
0 μg/mL sample

Calibration Mean Values


Calibration curves

Images of Our PCR Negative and Positive Controls

Negative control PCR sample
Positive Control PCR sample


PCR Results: PCR concentrations solved


PCR Results: Summary

  • Our positive control PCR result was 11.857 μg/mL
  • Our negative control PCR result was 3.544 μg/mL


Observed results

  • Patient 27929 : When this patient's DNA was exposed to the SYBR green solution, the SYBR green solution made it fluoresce.
  • Patient 34835 : When this patient's DNA was exposed to the SYBR green solution, the drop didn't fluoresce at all.


Conclusions

  • Patient 27929 : positive, because the DNA concentration values were greater than that of the positive control.
  • Patient 34835 : negative, because the DNA concentration values were less than that of the positive control.