Photo of the Single-Drop Fluorimeter Device. (Image used from Google Images)
SYBR Green Dye
SYBR Green Dye is a type of dye that is used to highlight when dsDNA is present because that is when it fluoresces the strongest. The dye fluoresces weak with water or single strands of DNA.
Single-Drop Fluorimeter
The Single-Drop Fluorimeter (as pictured above) is a custom device designed for the purpose of finding when a double-stranded piece of DNA is present. It fluoresces double-stranded DNA fragments in a green color, and does nothing to a solution without the double-stranded piece of DNA.
How the Fluorescence Technique Works
The process had several different variables that were chosen for very specific reasons. The slide to be used was chosen because it had a rough surface with glass circles exposed in it, causing the solution to concentrate at those locations and be easier to examine. The DNA dye chosen for the experiment was used because it was not completely soluble, which allowed for the light to glow instead of causing a reflective light off of the drop.
Procedure
Smart Phone Camera Settings
Type of smart phone used and adjusted camera settings.
Phone: HTC Droid Incredible 2
Flash: None
ISO setting: 800
White Balance: Auto
Exposure: +2
Saturation: +2
Contrast:-2
Calibration
Photo of how camera is pointing at, and focusing in on, the drop. (Image used from Google Images)
The phone was set up in the phone stand so that the camera was focusing on the drop. It was zoomed in far enough to notice the color of the drop and how it changed with each concentraton change.
Distance between the smart phone cradle and drop = 3.7 cm
Solutions Used for Calibration[See worksheet page 5]Image J Values for All Samples[See worksheet page 5]