BME100 s2016:Group6 W1030AM L5
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OUR TEAM
LAB 5 WRITE-UPPCR Reaction ReportOur team was successful in pipetting liquids during the PCR reaction. The team was able to properly transfer the DNA samples, nucleotides, enzymes, and primers after reviewing the pre-lab reading. The first stop on the pipettor is used to collect the exact measurement of liquid indicated on the pipettor. The second stop is used to completely release all the liquid from the pipettor, ensuring that the experiment is conducted accurately. Fluorimeter ProcedureWeb camera set-up After setting the timer on the iPhone to 3 seconds, the iPhone was laid horizontally in front of the fluorimeter. The iPhone was placed on a small platform to be able to image the drops and part of the glass slip the drops are on.
Data Collection and AnalysisImages of High, Low, and Zero Calf Thymus DNA zero dna: .5 DNA:
Calibrator Mean Values
Negative Positive PCR Results: PCR concentrations solved
PCR Results: Summary
In Patient 78117, the images show the appearance of DNA as the concentration becomes greater and the SYBR Green is more noticeable as it binds to the DNA. The PCR concentrations were diluted by 12 microLiters and had consistent results in the visibility of the DNA in the diluted solution.
For Patient 38765, the images have more drastic changes in the visibility of DNA as the PCR concentrations before dilution increase by larger increments. This increase in concentration of PCR is seen in the results of the Diluted Concentrations as they have proportional results. The SYBR Green binds with the DNA structure to give it a green visible look when fluorescent light is shined through the bead of solution. Conclusions
This patient compared to the positive control value does not line up with the images and results, yet is very close to the negative control value. When observing the images, this patient not only had darker visualizations of the DNA but this correlated to the control test for the negative control value image. The results back this up in that the original non-diluted concentration of PCR is within a half of a microliter of the control value.
Patient 2 is much closer to the positive control value because the concentration of DNA in the PCR Products both before dilution and after is much higher. The images representing patient 2 are closer in comparison with the images from the positive control value. Due to the increase in PCR concentration, the green light is more vibrant from the fluorescent light shining through the bead of solution. Patient 2 is closest in comparison to the values of the negative control.
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