BME100 f2016:Group7 W1030AM L5

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OUR TEAM

Name: Koop Bills
Name: Tyler Lent
Name: Maria Hanna
Name: Omar Maranon
Name: Israel Zaldivar


LAB 5 WRITE-UP

PCR Reaction Report

Overall the pipetting was fairly straight forward. Luckily, enough of us had worked with a micropipette before and as such the entire process went very smooth. The pre-lab was more or less helpful, maybe not entirely needed, but it did serve it's purpose in ensuring that we were able to get our results transferred as efficiently as possible. Yes we understood the difference (gotta say that it's a pretty nifty feature to have on there. I believe that we were able to successfully transfer over all available liquid, and no we did not have to change our labeling scheme!

Fluorimeter Procedure

Imaging set-up

The first thing we did was to place a holding apparatus (what could have been a cellphone stand) onto the upper counter top on the lab bench. We then proceeded to place in a cellphone to be used as the recording device for the lab. After ensuring that the cellphone was properly placed, stable, and after measuring the distance from a test drop on the fluorimeter, we then collected some standard images to use as a calibration curve.


Placing Samples onto the Fluorimeter

  1. Place the required amount of sample (80 micorliters) onto the fluorimeter]
  2. Place an equal amount of syber green on to the preexisting drop
  3. Ready the timer on the camera
  4. Close the lid to the "chamber" and take a total of 3 pictures
  5. Repeat steps 1-4 until you have captured an image of each necessary sample


Data Collection and Analysis

Images of High, Low, and Zero Calf Thymus DNA

5mgL.png .5mgL.png 0mgL.png


Calibrator Mean Values


Calibrator mean values.png


Calibration curves

Calibration curve 1.png Calibration curve 2.png

Images of Our PCR Negative and Positive Controls

Negative sample11.png Positive sample22.png

PCR Results: PCR concentrations solved

Pcr concentrations.png


PCR Results: Summary

  • Our positive control PCR result was 666.795616 μg/mL
  • Our negative control PCR result was 257.431264 μg/mL


Observed results

  • Patient 89545 : The image for this patient showed zero trace of green fluorescence. We took an average value for our pcr calculation of 36.130741333 μg/mL
  • Patient 25254 : This patient fluoresced a mild neon green color and the value we took from the chart was the one that "made sense" that being, 173.94976

μg/mL


Conclusions

  • Patient 89545 : with a value of 36.130741333 we can say that this patient is negative as the negative threshold is 257.431264
  • Patient 25254 : with a value of 173.94976 we can say that this patient is also negative (going to be honest and say that that doesn't seem right) as once again the threshold for a negative reading is 257.431264.