BME100 f2015:Group6 8amL5

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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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Contents

OUR TEAM

Name: Zach Humphreys
Name: Zach Humphreys
Name: Kianna Browning
Name: Kianna Browning
Name: Cameron McAllister
Name: Cameron McAllister
Name: Blake Browning
Name: Blake Browning
Name: Colten Hedrick
Name: Colten Hedrick
Name: Anuj Agrawal
Name: Anuj Agrawal


LAB 5 WRITE-UP

PCR Reaction Report

We used the pipettor to transfer the PCR reaction mix, patients 1 and 2's replicate mix, the positive DNa/primer mix, and the negative control. The key to this was proper technique. When drawing fluids, only push the pipettor to the first stop, but when ejecting fluids, push all the way to the second stop. Failure to do either of these will result in a difference in the actual amount as compared to the desired amount of substance. Next, we placed the tubes with the substances into the OpenPCR device.

Fluorimeter Procedure

Smart Phone Camera Settings

  • Type of Smartphone: iPhone 6S
    • Flash: Off
    • ISO setting: Auto
    • White Balance: Auto
    • Exposure: Auto
    • Saturation: Auto
    • Contrast: Auto


Camera set-up

  • Distance between the smart phone cradle and drop =


Placing Samples onto the Fluorimeter Image:Fluorimetergroup6.png

  1. Set volume on pipettor to 80 microliters.
  2. Place slide into fluorimeter such that the holes that will be used are aligned with the blue LED light.
  3. Firmly attach tip to pipettor.
  4. Pipette 80 microliters of SYBR Green I onto one of the middle holes of the slide.
  5. Eject pipette tip into waste basket.
  6. Firmly attach another tip.
  7. Pipette 80 microliters of .25 concentration of the calf thymus into the hole right above the SYBR Green I.
  8. Use timer on camera to take pictures of the solution in the dark under the blue LED light.
  9. Dispose of the liquid on the slide into the liquid waste basket.
  10. Repeat steps for the .5, 1, 2, and 5 concentrations of the calf thymus.


Data Collection and Analysis

Images of High, Low, and Zero Calf Thymus DNA


Calibrator Mean Values

Image: Tble_t2.png



Calibration curves
image: DOT_PLOT_1A_C.png Image: DOT_PLOT_2_CC.png


Images of Our PCR Negative and Positive Controls


PCR Results: PCR concentrations solved

Image: Table_table_table_ll.png


PCR Results: Summary

  • Our positive control PCR result was 3986962.3 μg/mL
  • Our negative control PCR result was 2182675.3 μg/mL


Observed results

  • Patient 78778:
  • Patient 63691:


Conclusions

  • Patient 78778: positive because values above positive control
  • Patient 63691: positive because values above positive control



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