BME100 f2013:W900 Group9 L5: Difference between revisions
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| [[Image:AllisonLind.jpg|100px|thumb|Name: [[User: Allison Lind | Allison Lind ]]]] | | [[Image:AllisonLind.jpg|100px|thumb|Name: [[User: Allison Lind | Allison Lind ]]]] | ||
| [[Image:Slankford_BME100.jpg|100px|thumb|Name: [[User: Sydney M. Lankford | Sydney M. Lankford ]] | | [[Image:Slankford_BME100.jpg|100px|thumb|Name: [[User: Sydney M. Lankford | Sydney M. Lankford ]]]] | ||
| [[Image:Kjw_528_-1600x1200-.jpg|100px|thumb|Name: [[User:Neel J. Patel | Neel J. Patel]] | | [[Image:Kjw_528_-1600x1200-.jpg|100px|thumb|Name: [[User:Neel J. Patel | Neel J. Patel]]]] | ||
| [[Image:Callie LaMarche. | | [[Image:Callie LaMarche.JPG|100px|thumb|Name: [[User: Callie M. Lamarche | Callie M. LaMarche ]]]] | ||
| [[Image:Julianna A. Brunner.jpg|100px|thumb|Name: [[User:Julianna A. Brunner | Julianna A. Brunner]] | | [[Image:Julianna A. Brunner.jpg|100px|thumb|Name: [[User:Julianna A. Brunner | Julianna A. Brunner]]]] | ||
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'''Single-Drop Fluorimeter'''<br> | '''Single-Drop Fluorimeter'''<br> | ||
A single-drop fluorimeter device detects PCR-amplified DNA molecules based on the amount of flourescent material in the sample. A blue LED light is shined upon the sample; a camera can then capture green light flourescened by the DNA. | A single-drop fluorimeter device detects PCR-amplified DNA molecules based on the amount of flourescent material in the sample. A blue LED light is shined upon the sample; a camera can then capture green light flourescened by the DNA. | ||
<br> | |||
[[Image:Group9_Fluorimeter.jpg|450px]] | |||
'''How the Fluorescence Technique Works'''<br> | '''How the Fluorescence Technique Works'''<br> | ||
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<br> | <br> | ||
==Procedure== | ==Procedure== | ||
'''Smart Phone Camera Settings'''<br> | '''Smart Phone Camera Settings'''<br> | ||
* Type of Smartphone: iPhone 5 | * Type of Smartphone: iPhone 5 | ||
** Flash: off | ** Flash: off | ||
** ISO setting: auto ( | ** ISO setting: auto (no option on model used) | ||
** White Balance: auto ( | ** White Balance: auto (no option on model used) | ||
** Exposure: auto ( | ** Exposure: auto (no option on model used) | ||
** Saturation: auto ( | ** Saturation: auto (no option on model used) | ||
** Contrast: auto ( | ** Contrast: auto (no option on model used) | ||
'''Calibration'''<br> | '''Calibration'''<br> | ||
The phone was placed in the cradle at an appropriate angle to view the slide from the side. The fluorimeter was raised in front of the phone to get the right angle (as if it phone were at eye-level with the slide). The camera was placed 7.5 cm away from the drop to ensure that the drop would be in focus. A light box was placed over the set up to provide darkness to take the picture of the light flowing through the DNA and SYBR Green 1. The fluorescence in the sample was easily seen with higher concentrations of SYBR Green 1. | |||
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The phone was placed in the cradle at an appropriate angle to view the slide from the side. The fluorimeter was raised in front of the phone to get the right angle. A light box was placed over the set up to provide darkness to take the picture of the light flowing through the DNA and SYBR Green 1. The fluorescence in the sample was easily seen with higher concentrations of SYBR Green 1. | [[Image:Group9_CameraSetup.jpg|450px]] | ||
* Distance between the smart phone cradle and drop = 7.5 cm | * Distance between the smart phone cradle and drop = 7.5 cm | ||
'''Solutions Used for Calibration' | '''Solutions Used for Calibration''' | ||
{| {{table}} width=700 | {| {{table}} width=700 | ||
|- | |- | ||
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'''Placing Samples onto the Fluorimeter''' | '''Placing Samples onto the Fluorimeter''' | ||
# | # Attach fresh tip to micropipette and set micropipette to 50 micro-liters | ||
# | # Push micropipette plunger to first stop, and place tip into SYBR Green 1 dye, release plunger | ||
# | # Lift tip out of SYBR Green 1 dye and hover micropipette tip directly between two holes of the fluorimeter slide | ||
# | # Push plunger down to second stop, allowing dye to release onto the slide | ||
# Repeat steps 1-4 but now with the DNA | |||
# After observing the solution on the fluorimeter, discard the solution | |||
# Push micropipette plunger to first stop, place tip in the solution on the slide, and release plunger | |||
# Repeat step 7 if solution still remains on slide, discard micropipette tip | |||
# Repeat steps 1-8 for remaining DNA, make sure to use each hole on the slide only once | |||
<br> | <br> | ||
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'''Representative Images of Samples''' | '''Representative Images of Samples''' | ||
'' | '''0.0 μg/mL DNA Concentration Image J Analysis''' | ||
<br><br> | |||
[[Image:0.0 DNA and Data.png|550px]] | |||
'''2.5 DNA Concentration Image J Analysis'''<br> | '''2.5 μg/mL DNA Concentration Image J Analysis''' | ||
[[Image:Group9_2.5ImageJScreenshot.jpg| | <br><br> | ||
[[Image:Group9_2.5ImageJScreenshot.jpg|550px]] | |||
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{| | {| | ||
| align="center" style="background:#f0f0f0;"|'''Calf Thymus DNA Concentration (FINAL), μg/mL''' | | align="center" style="background:#f0f0f0;"|'''Calf Thymus DNA Concentration (FINAL), μg/mL''' | ||
| align="center" style="background:#f0f0f0;"|'''''' | | align="center" style="background:#f0f0f0;"|'''Image Number''' | ||
| align="center" style="background:#f0f0f0;"|'''AREA''' | | align="center" style="background:#f0f0f0;"|'''AREA''' | ||
| align="center" style="background:#f0f0f0;"|'''Mean Pixel Value''' | | align="center" style="background:#f0f0f0;"|'''Mean Pixel Value''' | ||
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'''Fitting a Straight Line'''<br> | '''Fitting a Straight Line'''<br> | ||
[[Image:Group9_CalibrationCurve.jpg|500px]] | |||
<br> | <br> | ||
<!-- Note: Be sure to delete the text in brackets: ''[ ]'' --> | <!-- Note: Be sure to delete the text in brackets: ''[ ]'' --> |
Latest revision as of 23:44, 12 November 2013
BME 100 Fall 2013 | Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
OUR TEAM
LAB 5 WRITE-UPBackground InformationSYBR Green Dye Single-Drop Fluorimeter How the Fluorescence Technique Works
ProcedureSmart Phone Camera Settings
Solutions Used for Calibration
Data AnalysisRepresentative Images of Samples 0.0 μg/mL DNA Concentration Image J Analysis
Image J Values for All Samples
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