User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/25

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(FCS Measurements)
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**500pM DNA/600pM MB
**500pM DNA/600pM MB
**100pM DNA/120pM MB
**100pM DNA/120pM MB
 +
**500pM Oligo D fluorescent dye
 +
**100pM Oligo D fluorescent dye
 +
====Results====
 +
Oligo D Fluorescent Dye<br.>
 +
[[Image:FCS_data_2013_0625_Oligo_D_comparison_graphs.PNG]]<br.>
 +
<br.>
 +
Calibration Graph<br.>
 +
[[Image:FCS_data_2013_0625_Oligo_D_calibration.png]]<br.>
 +
<br.>
 +
Varied Concentrations of DNA and MB<br.>
 +
[[Image:FCS data 2013_0625 DNA-MB comparison graphs.PNG]]<br.>
 +
<br.>
 +
Calibration Graph<br.>
 +
[[Image:FCS data 2013_0625 DNA-MB calibration.png]]<br.>
 +
===Notes===
 +
From the calibration graph for the Oligo D, there is a concentration drop-off between 50 and 100pM.  More samples between this interval could be made to determine at what exact concentration the limit occurs.  Otherwise, the trend between concentration of Oligo D and 1/<N> value is linear. The graphs were consistent with the exception of the 50pM sample, which could be due to the concentration drop-off observed in the calibration graph.<br.>
 +
From the calibration graph for the DNA/MB samples, a concentration limit was not reached.  This means samples can be further diluted and run.  The graphs were consistent with the exception of the final sample, 100pM DNA/120pM MB, which was expected to have a higher y-intercept value. This may be a concentration limit similar to the Oligo D, but was not evident in the <N> value.  Samples will be made and run again tomorrow.<br.>
 +

Revision as of 15:37, 25 June 2013

FCS Data Main project page
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June 25, 2013

FCS Measurements

  • Samples prepared to be taken up to run on the FCS:
    • Antibodies with Alexa 532
    • Green fluorescent beads diluted 100x
    • Green fluorescent beads diluted 1000x
    • 100nM DNA/120nM MB
    • 1nM DNA/1.2nM MB
    • 500pM DNA/600pM MB
    • 100pM DNA/120pM MB
    • 500pM Oligo D fluorescent dye
    • 100pM Oligo D fluorescent dye

Results

Oligo D Fluorescent Dye
Image:FCS_data_2013_0625_Oligo_D_comparison_graphs.PNG

Calibration Graph
Image:FCS_data_2013_0625_Oligo_D_calibration.png

Varied Concentrations of DNA and MB
Image:FCS data 2013_0625 DNA-MB comparison graphs.PNG

Calibration Graph
Image:FCS data 2013_0625 DNA-MB calibration.png

Notes

From the calibration graph for the Oligo D, there is a concentration drop-off between 50 and 100pM. More samples between this interval could be made to determine at what exact concentration the limit occurs. Otherwise, the trend between concentration of Oligo D and 1/<N> value is linear. The graphs were consistent with the exception of the 50pM sample, which could be due to the concentration drop-off observed in the calibration graph.
From the calibration graph for the DNA/MB samples, a concentration limit was not reached. This means samples can be further diluted and run. The graphs were consistent with the exception of the final sample, 100pM DNA/120pM MB, which was expected to have a higher y-intercept value. This may be a concentration limit similar to the Oligo D, but was not evident in the <N> value. Samples will be made and run again tomorrow.



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