BME100 s2014:T Group2 L5: Difference between revisions
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'''SYBR Green Dye'''<br> | '''SYBR Green Dye'''<br> | ||
''An asymmetrical cyanine dye that stains the nucleic acids of DNA. The dye binds to the DNA, turning it green in order to detect whether or not it's a DNA flourimeter. If a sample does not turn green in the flourimeter, then it does not contain DNA. | ''An asymmetrical cyanine dye that stains the nucleic acids of DNA. The dye binds to the DNA, turning it green in order to detect whether or not it's a DNA flourimeter. If a sample does not turn green in the flourimeter, then it does not contain DNA.'' | ||
[[Image:20140403_110450.jpg]] | |||
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'''Calibration'''<br> | '''Calibration'''<br> | ||
'' The smartphone was placed in the cradle perpendicular to the Fluorimeter. The cradle was measured at 12.5 cm away from the fluorimeter | '' The smartphone was placed in the cradle perpendicular to the Fluorimeter. The cradle was measured at 12.5 cm away from the fluorimeter.'' | ||
[[Image:20140403_110615.jpg]] | |||
* Distance between the smart phone cradle and drop = 12.5cm | |||
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| 0 || 80 || 80 || 0 | | 0 || 80 || 80 || 0 | ||
|} | |} | ||
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'''Representative Images of Samples''' | '''Representative Images of Samples''' | ||
Below is an image depicting a sample analyzed using imagej that had a concentration of 0. | |||
[[Image:0 oval.png]] | |||
This image shows a sample with concentration of 2.5, as is apparent there is a notable difference between the concentration of 0 as the green light is not visible in the picture above. | |||
[[Image:2.5 oval.png]] | |||
'''Image J Values for All Samples''' | '''Image J Values for All Samples''' | ||
{| | {| | ||
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| align="center" style="background:#f0f0f0;"|'''Corrected PCR Product Concentration, µg /mL''' | | align="center" style="background:#f0f0f0;"|'''Corrected PCR Product Concentration, µg /mL''' | ||
|- | |- | ||
| 1. Positive||14109839||1.210984||0.100915325 | |||
| 1. | |||
|- | |- | ||
| 2. A1||5644840||0.364484||0.030373667 | | 2. A1||5644840||0.364484||0.030373667 | ||
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| 4. A3||5443461||0.344346||0.028695508 | | 4. A3||5443461||0.344346||0.028695508 | ||
|- | |- | ||
| 5. | | 5. Negative||6025545||0.402555||0.033546208 | ||
|- | |- | ||
| 6. B1||13694379||1.169438||0.097453158 | | 6. B1||13694379||1.169438||0.097453158 | ||
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|} | |} | ||
The values for concentrations and corrected concentrations can be found above. Note that the positive control possessed a higher concentration of DNA than the negative sample. This is good since those values are used as a threshold for determining if the others are either positive or negative. In the images the negative sample appeared to reflect little to no green at all whereas the positive was very brightly illuminated by the fluorescent light. | |||
Patient ID: 77842 Is listed above using the A values such that the range of concentrations were close to .03. This value and the striking resemblance color-wise to the negative sample leads one to believe that it did not possess the DNA that was tested. | |||
Patient ID:26204 Was labeled using the B values above and was found to have a much higher concentration of the DNA sample. This sample appeared to have been very green when analyzing the images through the analysis software. Given this it makes since to infer that the sample was tested positive for the DNA. | |||
'''Fitting a Straight Line'''<br> | '''Fitting a Straight Line'''<br> | ||
Below is the graph illustrating the corrected concentrations compared to the intden values found through imagej. | |||
[[Image:2graphbmepcr.jpg]] | |||
<br> | <br> |
Latest revision as of 20:33, 16 April 2014
BME 100 Spring 2014 | 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
ProcedureSmart Phone Camera Settings
The smartphone was placed in the cradle perpendicular to the Fluorimeter. The cradle was measured at 12.5 cm away from the fluorimeter.
Solutions Used for Calibration [Instructions: See worksheet page 6.]
Placing Samples onto the Fluorimeter
Data AnalysisRepresentative Images of Samples Below is an image depicting a sample analyzed using imagej that had a concentration of 0.
The values for concentrations and corrected concentrations can be found above. Note that the positive control possessed a higher concentration of DNA than the negative sample. This is good since those values are used as a threshold for determining if the others are either positive or negative. In the images the negative sample appeared to reflect little to no green at all whereas the positive was very brightly illuminated by the fluorescent light. Patient ID: 77842 Is listed above using the A values such that the range of concentrations were close to .03. This value and the striking resemblance color-wise to the negative sample leads one to believe that it did not possess the DNA that was tested. Patient ID:26204 Was labeled using the B values above and was found to have a much higher concentration of the DNA sample. This sample appeared to have been very green when analyzing the images through the analysis software. Given this it makes since to infer that the sample was tested positive for the DNA. Fitting a Straight Line
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