User:Allison K. Alix/Notebook/Thesis Research/2013/11/05

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Current revision (16:08, 5 November 2013) (view source)
 
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> November 5th, 2013</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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[[Image:OWW_AUNP_DNA_pic.png]]
[[Image:OWW_AUNP_DNA_pic.png]]
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==Calculating AuNP/DNA Concentration==
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AuNP: abs @ 532 = 0.03
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ε = 92,382,878
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c= A/εb
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c = 3.2473x10<sup>-10</sup>M = 0.325nM
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DNA: abs @ 254 = 0.399
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ε = 607,500
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c = 6.5679E-07M = 656.6nM
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==Addition of MB==
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1) Add 6uL 7.61uM MB to 50uL AuNP/DNA Solution
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NEW Concentrations:
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MB: 815nM
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DNA: 586nM
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AuNP: 0.29nM
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2) Heat for 25 min at ~76.0°C
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3) Run FCS Measurements. Begin with pure sample. Because the AuNP concentration is so low (290pM) it might be hard to get signal from anything lower.
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==Other Notes==
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The UV-Vis spectra from the beacon alone did not yield the expected results. Not only were the spectra out of order (expecting to see highest concentration have the highest absorbance) but it was jagged, and not smooth like ti normally is. This might be attributed to the fact that the samples were not freshly made (prepared 10/30) and the DNA could have either clumped together causing inaccurate scattering patterns of light or settled to the bottom. (see image below)
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[[Image:OWW_MB_uv.png]]
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Current revision

November 5th, 2013 Main project page
Previous entry      Next entry

To Do

  • Run UV-Vis of MB samples to determine concentraiton
  • Add MB to AuNP/DNA Sample. Run FCS.

Data

Image:OWW_AUNP_DNA_pic.png

Calculating AuNP/DNA Concentration

AuNP: abs @ 532 = 0.03

ε = 92,382,878

c= A/εb

c = 3.2473x10-10M = 0.325nM

DNA: abs @ 254 = 0.399

ε = 607,500

c = 6.5679E-07M = 656.6nM

Addition of MB

1) Add 6uL 7.61uM MB to 50uL AuNP/DNA Solution

NEW Concentrations:

MB: 815nM

DNA: 586nM

AuNP: 0.29nM

2) Heat for 25 min at ~76.0°C

3) Run FCS Measurements. Begin with pure sample. Because the AuNP concentration is so low (290pM) it might be hard to get signal from anything lower.

Other Notes

The UV-Vis spectra from the beacon alone did not yield the expected results. Not only were the spectra out of order (expecting to see highest concentration have the highest absorbance) but it was jagged, and not smooth like ti normally is. This might be attributed to the fact that the samples were not freshly made (prepared 10/30) and the DNA could have either clumped together causing inaccurate scattering patterns of light or settled to the bottom. (see image below)

Image:OWW_MB_uv.png


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