OUR TEAM
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LAB 5 WRITE-UP
Procedure
Smart Phone Camera Settings
- Type of Smartphone:
- Flash: NO Flash
- ISO setting:800
- White Balance: Auto
- Exposure: Zero
- Saturation:Standard
- Contrast:Auto
Calibration
- Distance between the smart phone cradle and drop = 6.5 cm
Solutions Used for Calibration
Init. Conc. 2X Calf Thymus DNA (micrograms/mL) |
Vol. 2X DNA solution (micro-mL) |
Vol. SYBR GREEN 1 DYE (micro-mL) |
Final DNA Concentration STBR GREEN 1 Solution (Micrograms/mL)
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5 |
80 |
80 |
row 2 cell 4
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2 |
80 |
80 |
row 3 cell 4
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1 |
80 |
80 |
row 1 cell 4
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0.5 |
80 |
80 |
row 2 cell 4
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0.25 |
80 |
80 |
row 3 cell 4
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0 |
80 |
80 |
row 3 cell 4
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Placing Samples onto the Fluorimeter
- Place fluorimeter on the table
- Place slide in the fluorimeter with the smooth side down hydrophobic side up
- Adjust slide to the light shines between the dots on the slide
- Place 80 micro-mL of SYBR green 1 on slide between the middle dots
- Place 80 micro-mL of calibration on the sphere you already placed to a 160 micro-mL shpere
- Adjust the shpere placement for the light and phone distance in the holder
- Record Iphone distance
- Cover the assembly and make small adjustments
- Take a photo on delay and cover
- Send photo to be recorded for ImageJ analysis
- Remove the 160 micro-mL
- Discard the slide
- Repeat steps 1-12
Data Analysis
Representative Images of Negative and Positive Samples
Image J Values for All Calibrator Samples
EXCEL Table1 RAW DATA
Final DNA Concentration in SYBR Green 1 solution (microgram/mL) |
Area |
Mean Pixel Value |
RAWINTDEN OF THE DROP |
RAWINTDEN OF THE BACKGROUND
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0 |
Cell |
Cell |
Cell |
Cell
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0.25 |
Cell |
Cell |
Cell |
Cell
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0.5 |
Cell |
Cell |
Cell |
Cell
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1 |
Cell |
Cell |
Cell |
Cell
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2 |
Cell |
Cell |
Cell |
Cell
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N |
Cell |
Cell |
Cell |
Cell
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P |
Cell |
Cell |
Cell |
Cell
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1.1 |
Cell |
Cell |
Cell |
Cell
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1.2 |
Cell |
Cell |
Cell |
Cell
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1.3 |
Cell |
Cell |
Cell |
Cell
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2.1 |
Cell |
Cell |
Cell |
Cell
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2.2 |
Cell |
Cell |
Cell |
Cell
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2.3 |
Cell |
Cell |
Cell |
Cell
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EXCEL Table 2 Edited to included background-subtractions
Final DNA Concentration in SYBR Green 1 solution (microgram/mL) |
Area |
Mean Pixel Value |
RAWINTDEN OF THE DROP |
RAWINTDEN OF THE BACKGROUND |
RAWINTDEN-DROP Background
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Cell |
Cell |
Cell |
Cell |
Cell |
Cell
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Cell |
Cell |
Cell |
Cell |
Cell |
Cell
|
Cell |
Cell |
Cell |
Cell |
Cell |
Cell
|
Cell |
Cell |
Cell |
Cell |
Cell |
Cell
|
Cell |
Cell |
Cell |
Cell |
Cell |
Cell
|
Cell |
Cell |
Cell |
Cell |
Cell |
Cell
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Calibration curve
PCR Results Summary
- Our positive control PCR result was ____ μg/mL
- Our negative control PCR result was ____ μg/mL
Observed results
- Patient _____ :
- Patient _____ :
Conclusions
- Patient _____ :
- Patient _____ :
SNP Information & Primer Design
Background: About the Disease SNP
Primer Design and Testing
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