User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/18

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(Autocreate 2013/06/18 Entry for User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence)
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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;"> Replication Attempts</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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==Entry title==
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==June 18, 2013==
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* Insert content here...
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===FCS===
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*Calibration:
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**Alexa 532 Antibodies at 500pM and 100pM
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**green fluorescent beads diluted 10000x
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*Samples:
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**1-8 from 06/13/13
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No fluorescence obvserved.
 +
===Sample Prep: Remake 727.5nM DNA/9.7nM AuNPs===
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*Making 100mM DTT in 4% TEA
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*Extraction procedure with ethyl acetate
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*Combine equal volumes of 19.4nM AuNPs with 1455nM DNA from extraction to make 727.5nM DNA/9.4nM AuNP solution
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*Centrifuge four times, collecting four supernatants,at 4000rpm for 5 min intervals then re-dispersing in 50mM HEPES buffer
 +
*Take absorbance spectra for concentrated sample and four supernatants
 +
===Sample Prep: Remake 727.5nM DNA and 14.835nM AuNP Solutions===
 +
*Take fluorescence spectra, exciting at 532nm for the Molecular Beacon data
 +
===Sample Prep: Additional DNA Concentrations for DNA/ThT Correlation Graph===
 +
*Make the following solutions (use PBS for dilutions):
 +
**800nM DNA/2.5μM ThT
 +
**1091.25nM DNA/2.5μM ThT
 +
**1200nM DNA/2.5μM ThT
 +
===Absorbance and Fluorescence Data===
 +
*Absorbance data taken for DNA/ThT samples, (needs to be rerun due to inconsistencies)and extracted AuNPs with DNA and four supernatants
 +
**Samples might be remade before re-running
 +
*Fluorescence data taken for DNA/ThT samples, DNA and ThT comparison
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Current revision

Replication Attempts Main project page
Previous entry      Next entry

June 18, 2013

FCS

  • Calibration:
    • Alexa 532 Antibodies at 500pM and 100pM
    • green fluorescent beads diluted 10000x
  • Samples:
    • 1-8 from 06/13/13

No fluorescence obvserved.

Sample Prep: Remake 727.5nM DNA/9.7nM AuNPs

  • Making 100mM DTT in 4% TEA
  • Extraction procedure with ethyl acetate
  • Combine equal volumes of 19.4nM AuNPs with 1455nM DNA from extraction to make 727.5nM DNA/9.4nM AuNP solution
  • Centrifuge four times, collecting four supernatants,at 4000rpm for 5 min intervals then re-dispersing in 50mM HEPES buffer
  • Take absorbance spectra for concentrated sample and four supernatants

Sample Prep: Remake 727.5nM DNA and 14.835nM AuNP Solutions

  • Take fluorescence spectra, exciting at 532nm for the Molecular Beacon data

Sample Prep: Additional DNA Concentrations for DNA/ThT Correlation Graph

  • Make the following solutions (use PBS for dilutions):
    • 800nM DNA/2.5μM ThT
    • 1091.25nM DNA/2.5μM ThT
    • 1200nM DNA/2.5μM ThT

Absorbance and Fluorescence Data

  • Absorbance data taken for DNA/ThT samples, (needs to be rerun due to inconsistencies)and extracted AuNPs with DNA and four supernatants
    • Samples might be remade before re-running
  • Fluorescence data taken for DNA/ThT samples, DNA and ThT comparison


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