SYBR Green Dye [SBYR green dye is a nucleic acid-sensitive gel dye that is useful in pinpointing double stranded DNA in samples within open PCR machines and other utilities such as the single-drop fluorimeter.]
Single-Drop Fluorimeter [The single-drop fluorimeter is a fluorescence-based DNA detection device that aids in bringing to light PCR-Amplified DNA. The DNA sample is placed on a plate placed upon the machine and shined with a blue LED light. When shone upon, the sample's DNA begins to emit a green light; able to be captured with a cell phone camera.]
How the Fluorescence Technique Works [In your own words, a summary of the information from page 9 of the worksheet]
Procedure
Smart Phone Camera Settings
Iphone 5
Flash: None
ISO setting: NA
White Balance: NA
Exposure: NA
Saturation: NA
Contrast: NA
Calibration
Photo of Fluorimeter
Distance between the smart phone cradle and drop = 8cm
Solutions Used for Calibration
Calf Thymus DNA solution concentration (microg/mL)
Volume of the 2X DNA solution (uL)
Volume of the SYBR GREEN I Dye Solution (uL)
Final DNA concentration in PicoGreen Assay (ng/mL)
0
80
80
blank
.25
80
80
.125
.5
80
80
.25
1
80
80
.5
2
80
80
1
5
80
80
2.5
Placing Samples onto the Fluorimeter
Get Slide, Micropipette and vial of sample
Place slide into fluorimeter
Take 80 micro liters of sample into micropipette
Place 4 drops onto the slide, slide the slide over to where the light hits, don't use the same place twice (place new drops in different area)
Make sure the drops form a ball and don't spread around individually
Clean Up Workstation
Data Analysis
Representative Images of Samples
[Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with no DNA]
[Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with DNA (positive signal)]
Image J Values for All Samples[See worksheet page 5]
row 1 cell 1
row 1 cell 2
row 1 cell 3
row 1 cell 4
row 1 cell 5
row 2 cell 1
row 2 cell 2
row 2 cell 3
row 2 cell 4
row 2 cell 5
row 3 cell 1
row 3 cell 2
row 3 cell 3
row 3 cell 4
row 2 cell 5
[Add more rows as needed]
Fitting a Straight Line [Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 8]