Difference between revisions of "User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/03"

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Measurements and Sample Prep for FCS</span>
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Measurements and Sample Prep for FCS</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>}}
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*ThT- 2.2245μM
 
*ThT- 2.2245μM
 
<br.>
 
<br.>
=====Absorbance and Fluorescence Data=====
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=====Data=====
 
[[Image:Abs_data_thiol_DNA,_AuNP,_MB_06032013.png]]
 
[[Image:Abs_data_thiol_DNA,_AuNP,_MB_06032013.png]]
 
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[[Image:Fluor_data_thiol_DNA,_AuNP,_ThT_06032013.png]]
 
[[Image:Fluor_data_thiol_DNA,_AuNP,_ThT_06032013.png]]
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====Notes====
 
====Notes====
 
Samples containing Molecular Beacon excited at 532nm<br.>
 
Samples containing Molecular Beacon excited at 532nm<br.>

Latest revision as of 22:43, 26 September 2017

Owwnotebook icon.png Measurements and Sample Prep for FCS Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

June 3, 2013

Absorbance and Fluorescence Data

Finish preparing thiol DNA, AuNP and ThT sample

2[ThT]:1[DNA]→ 2.2245μM ThT<br.> 10% of 250μL=25μL (within 10% allows for accurate concentration comparison)<br.> Total solution volume: 275μL<br.> (2.2245×10-6M)(275×10-6L)=6.11738×10-10moles ThT<br.> (6.11738×10-10moles)/(25×10-6L)=24.5μM ThT<br.> <br.> To make 24.5μM ThT:<br.> (120μM)V=(24.5μM)(25μL)<br.> V=5.1μL in 19.9μL PBS buffer<br.> <br.> Add 25μL of 24.5μM ThT to 250μL DNA/AuNP solution<br.> New concentrations:

  • thiol DNA- 1.011μM
  • AuNP- 13.486μM
  • ThT- 2.2245μM

<br.>

Data

Abs data thiol DNA, AuNP, MB 06032013.png <br.> <br.> Abs data thiol DNA, ThT 06032013.png <br.> <br.> Fluor data thiol DNA, AuNP, MB 06032013.png <br.> <br.> Fluor data thiol DNA, AuNP, ThT 06032013.png

Notes

Samples containing Molecular Beacon excited at 532nm<br.> Samples containing ThT excited at 440nm<br.>

Sample Preparation for FCS and Calculations

Thiol DNA, MB, AuNP Solution Dilutions
  • Starting Concentrations:
    • AuNP- 9.03nM
    • MB-1268nM
    • Thiol DNA- 677.157nM

1. 1nM AuNPs<br.> (9.03nM)V=(1nM)(30μL)<br.> V=3.32μL in 26.68μL PBS buffer

  • New Concentrations:
    • AuNP- 1nM
    • MB- 140.33nM
    • Thiol DNA- 74.94nM

2. 500pM AuNPs<br.> 15μL 1nM solution in 15μL PBS buffer

  • New Concentrations:
    • AuNP- 500pM
    • MB- 70.165nM
    • Thiol DNA- 37.47nM

3. 100pM AuNPs<br.> 6μL 500pM solution in 24μL PBS buffer

  • New Concentrations:
    • AuNP- 100pM
    • MB- 14.033nM
    • Thiol DNA- 7.55nM
Thiol DNA, MB Solution Dilutions
  • Starting Concentrations:
    • MB- 1268nM
    • Thiol DNA- 727.5nM

4. 140.33nM MB<br.> 3.32μL in 26.68μL PBS buffer

  • New Concentrations:
    • MB- 140.33nM
    • 80.51nM

5. 70.165nM MB<br.> 15μL 140.33nM MB in 15μL PBS buffer

  • New Concentrations:
    • MB- 70.165nM
    • Thiol DNA- 40.26nM

6. 14.033nM MB<br.> 6μL 70.165nM MB in 24μL PBS buffer

  • New Concentrations:
    • MB- 14.033nM
    • Thiol DNA- 40.26nM
MB Solution Dilutions
  • Starting Concentrations:
    • MB- 1268nM

7. 140.33nM MB<br.> 3.32μL in 26.68μL PBS buffer<br.> <br.> 8. 70.165nM MB<br.> 15μL 140.33nM MB in 15μL PBS buffer<br.> <br.> 9. 14.033nM MB<br.> 6μL 70.165nM MB in 24μL PBS buffer<br.>