BME100 s2016:Group3 W1030AM L5

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Contents

OUR TEAM

Name: Mark Weser
Name: Mark Weser
Name: Ryan Wood
Name: Ryan Wood
Name: Morgan Murray
Name: Morgan Murray
Name: Kori Staples
Name: Kori Staples
Name: Makarios Begay
Name: Makarios Begay
Name: Lexi Nelson
Name: Lexi Nelson


LAB 5 WRITE-UP

PCR Reaction Report

Our team probably has extensive pipetting experience; we have over ten confirmed lab experiences with pipettes. Since we read the pre-lab reading in the lab, it became known as lab reading, and it was helpful. The first stop of the pipette will be utilized to measure for the amount while the second stop will be utilized to hold the solution until release.

Fluorimeter Procedure

Web camera set-up
The camera phone should be a device which can take photos using a timer. The webcam should be placed in the fluorimeter in such a fashion that the view is perpendicular relative to the drop. Thus, the view should be similar to the stereotypical view of looking down a long road. As a result, the camera phone should be able to stand by itself and take a photo. In addition, there are particular functions which must be changed in order to effectively capture a stellar image: the flash should be deactivated, the ISO set to 800 or higher, set white balance to auto, set exposure and saturation to the maximum, and contrast to the minimum.


Placing Samples onto the Fluorimeter
Before any thoughts about the placement of the samples occur, one should check that the placement of the smooth side of the slide is facing down. Once checked, the eighty micro-liter solution of SYBR Green 1 should be placed in the center of the first middle circles. Then, the eighty micro-liter solution of the given sample is placed on top of the SYBR Green 1 drop.


Data Collection and Analysis

Images of High, Low, and Zero Calf Thymus DNA







Calibrator Mean Values


Calibration curves


Images of Our PCR Negative and Positive Controls





Negative Picture

Positive Picture


PCR Results: PCR concentrations solved


PCR Results: Summary

  • Our positive control PCR result was 14.1219 μg/mL
  • Our negative control PCR result was 12.6547778 μg/mL


Observed results

  • Patient 35239 : This patient's samples were colorless compared to the green fluorescent positive control sample. The PCR concentrations were calculated to be around -16.2409μg/mL.
  • Patient 54995 : This patient served as the negative control, meaning the samples should be colorless. Our results showed that the patient's samples were in fact colorless and the PCR concentrations were calculated to be around -15.7526μg/mL.


Conclusions

  • Patient 35239 : This patient produced a negative result, because the values of all three trials differed from the positive control's threshold of -16.2409- -16.3709μg/mL.
  • Patient 54995 : This patient's PCR concentration levels (-15.7526μg/mL) were well below the positive control, so this patient also produced a negative result.



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