SYBR Green Dye [Instructions: A short summary describing SYBR green dye]
Single-Drop Fluorimeter [Instructions: A description of the single-drop fluorimeter device. Add a PHOTO for bonus points]
How the Fluorescence Technique Works [Instructions: In your own words]
Procedure
Smart Phone Camera Settings
Type of Smartphone: Apple iPhone 5S
Flash: Off
ISO setting: Auto
White Balance: Auto
Exposure: High
Saturation: High
Contrast: Low
Calibration
The camera was set up by placing the iPhone 5S in the holding compartment of the lab equipment. It was measured to be 6 centimeters away from the origin of the light emission. The settings (stated above) were adjusted; the height of the fluorimeter was adjusted in order to achieve a perfect side-angle perspective of the "beach ball" shaped liquid drop.
Distance between the smart phone cradle and drop = 8 centimeters
Solutions Used for Calibration
Initial Concentration of 2X Calf Thymus DNA solution (μg/mL)
Volume of the 2X DNA solution (μL)
Volume of the SYBR GREEN I
Final DNA concentration in SYBR Green I solution (μg/mL)
5
80
80
2.5
2
80
80
1.0
1
80
80
0.5
0.5
80
80
0.25
0.25
80
80
0.125
0
80
80
0
Placing Samples onto the Fluorimeter
Before placing samples onto the fluorimeter, turn on the Blue LED light & have the camera on the smartphone ready.
Insert the glass sampling surface, with the smooth side facing down.
Position the glass sampling surface so that the light emits between the first two rows of the glass.
Place an 80 μL drop of SYBR Green I in the middle of the first two rows.
Swap out the pipette tip with a new tip.
Place an 80 μL drop of a specifically concentrated Calf Thymus DNA solution onto the top of the previously placed SYBR Green I drop. Every three 160 μL sample drops will vary in concentrated Calf Thymus DNA solution.
In order to place a new sample drop, remove the 160 μL solution, move the glass sampling surface to the next two available rows, and repeat the process of placing two new 80 μL drops onto each other (as stated before).
Data Analysis
Representative Images of Samples
Sample w/o DNA
Sample w/ DNA
Image J Values for All Samples
Final DNA Concentration in SYBR Green I Solution (μL/mL)
RAWINTDEN of the Drop
RAWINTDEN of the Background
RAWINTDEN Drop - RAWINTDEN Background
0.5
4421613
187151
4234462
0.5
4521561
175412
4346149
0.5
4159878
179854
3980024
1.0
5904302
195454
5708848
1.0
6048248
187945
5860303
1.0
6248451
198454
6049997
2.5
8558810
212416
8346394
2.5
8821541
205451
8616090
2.5
8258495
215411
8043084
0.0
3201562
167842
3033720
0.0
3158472
154952
3003520
0.0
3584956
151231
3433725
0.125
1124652
165451
959201
0.125
1413586
167851
1245735
0.125
1248412
170245
1078167
0.25
3952152
168672
3783480
0.25
3754546
171541
3583005
0.25
4132142
172164
3959978
Fitting a Straight Line
[Instructions: Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 9]