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.
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
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Placing Samples onto the Fluorimeter
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.
Move the drop to focus the blue light so that it hits the black sensor on the opposite side.
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.
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.
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.
BME 100 Fall 2013
