BME100 f2013:W1200 Group15 L5

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OUR TEAM

Name: Patrick McFarland
Taylor Olvey
Name: Zac Roy
Name: Greg Berghost
Name:Tameem Jamal


LAB 5 WRITE-UP

Background Information

SYBR Green Dye

The SYBR green dye is a clear liquid that will luminate when a blue light shined upon it. The SYBR green dye is able to be mixed with other substances (DNA)that will glow under a blue light.


Single-Drop Fluorimeter

A glass plate is held down by the Fluorimeter with a blue light shining across the glass plate. On the glass plate will be your mixture which would include the DNA mixed with the SYBR green dye.
Picture.png

How the Fluorescence Technique Works
The Fluorescence technique works by the SYBR green dye luminating when a blue light shines on it. The SYBR green dye absorbs the blue light then reflects the green color that is seen on the glass plate.



Procedure

Smart Phone Camera Settings
[Instructions: The type of smart phone you used and how you adjusted the camera settings, if applicable (see worksheet page 4).]

  • Type of Smartphone: Galaxy-3X
    • Flash: N/A
    • ISO setting: 800
    • White Balance: auto
    • Exposure:+2.0
    • Saturation: N/A
    • Contrast: N/A

Calibration

  • Distance between the smart phone cradle and drop = 4.10cm


Solutions Used for Calibration

DNA solution (microg/ml) Volume of DNA solution (micro-Liters) Volume of SYBR Green solution (micro-Liters) Final Concentraition of DNA + SYBR (ng/mL)
5 80 80 2.5
2 80 80 1
1 80 80 0.5
0.05 80 80 0.25
0.25 80 80 0.125
0 80 80 blank


Placing Samples onto the Fluorimeter

  1. Use the micro pipette to place 80 microliters of SYBR GREEN I in between the first two circles on the the slide. Then use the pipette to add 80 microliters of the intended DNA solution to the SYBR GREEB I.
  2. Move the drop to focus the blue light so that it hits the black sensor on the opposite side.
  3. Use the light box when taking the two photos with the phone to block out interfering light. Be sure never to move the phone once it is in place.
  4. Take off the box and use the pipette to get the drop off the slide. Then move the slide to the next spot and set up the next sample.
  5. Be sure to do three tests of the same concentration with different drops on the same spot of the slide. Move onto the next concentration.


Data Analysis

Representative Images of Samples

0-1.jpg

5ml-1.jpg


Image J Values for All Samples


Fitting a Straight Line

Calfthymus123.jpg