BME100 f2014:Group1 L5

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BME 100 Fall 2014 Home
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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: Hawley Helmbrecht
Name: Timothy Black
Name: Sarah Fakhoury
Role(s)
Name: Prerna Gupta
Role(s)
Name: Jonathan Riecker
Name: Michael Pineda
Role(s)


LAB 5 WRITE-UP

Procedure

Smart Phone Camera Settings

  • Type of Smartphone: Samsung S3
    • Flash: No Flash
    • ISO setting: 800
    • White Balance: Auto
    • Exposure: +2.0
    • Saturation: No setting to affect Saturation
    • Contrast: Auto Contrast Off


Calibration

Place the phone in the cradle with the camera lens lined up so that it shows a view of the slide from the side (edge-on). Once the camera is set up in the cradle and lined up with the side of the slide, adjust the slide so that the light illuminates the center of the drop and the drop focuses light on the other side of it. After adjusting the slide itself, change the distance of the smartphone from the fluorimeter in order to keep the drop in focus but keep the view close.

  • Distance between the smart phone cradle and drop = 5 cm


Solutions Used for Calibration

PCR Product Label Tube Volume of the DILUTED PCR Product Solution (μL) Volume of the SYBR GREEN I Dye Solution (μL) Dilution 1 Dilution 2 Total Dilution (Simplified Fraction)
Positive 80 80 1/6 1/2 1/12
Negative 80 80 1/6 1/2 1/12
G1 1-1 80 80 1/6 1/2 1/12
G1 1-2 80 80 1/6 1/2 1/12
G1 1-3 80 80 1/6 1/2 1/12
G1 2-1 80 80 1/6 1/2 1/12
G1 2-2 80 80 1/6 1/2 1/12
G1 2-3 80 80 1/6 1/2 1/12



Placing Samples onto the Fluorimeter

  1. Find the rough side of the slide (making sure to only touch the slide while wearing gloves)
  2. Place the slide, with the rough side up, in the fluorimeter
  3. Locate the two clear circles in the middle of the slide and add 80 μL of SYBR Green I solution
  4. On the same circle with the SYBR Green I solution, place 80 μL of Sample/Calibration drops
  5. Adjust the slide so that the middle of the drop is illuminated by the light and the drop becomes focused on the side


Data Analysis

Representative Images of Negative and Positive Samples


Image J Values for All Calibrator Samples

Concentration of 2X Calf Thymus DNA solution(micrograms/mL) Volume of the 2X DNA solution (μL) Volume of the SYBR GREEN I Dye Solution (μL) Final DNA concentration in SYBR Green I solution (μg/mL)
5 80 80 2.5
2 80 80 1
1 80 80 0.5
0.5 80 80 0.25
0.25 80 80 0.125
0 80 80 0


Solutions Used for Calibration


Calibration curve


PCR Results Summary

  • Our positive control PCR result was ____ μg/mL
  • Our negative control PCR result was ____ μg/mL

Observed results

  • Patient _____ :
  • Patient _____ :

Conclusions

  • Patient _____ :
  • Patient _____ :




SNP Information & Primer Design

Background: About the Disease SNP


Primer Design and Testing

include a screen capture of the results web page