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

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(June 3, 2013)
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==June 3, 2013==
 
==June 3, 2013==
 
===Absorbance and Fluorescence Data===
 
===Absorbance and Fluorescence Data===
===Notes===
+
Finish preparing thiol DNA, AuNP and ThT sample<br.>
 +
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<sup>-6</sup>M)(275×10<sup>-6</sup>L)=6.11738×10<sup>-10</sup>moles ThT<br.>
 +
(6.11738×10<sup>-10</sup>moles)/(25×10<sup>-6</sup>L)=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.>
 +
[[Image:Abs_data_thiol_DNA,_AuNP,_MB_06032013.png]]
 +
<br.>
 +
<br.>
 +
[[Image:Abs_data_thiol_DNA%2C_ThT_06032013.png]]
 +
<br.>
 +
<br.>
 +
[[Image:Fluor_data_thiol_DNA,_AuNP,_MB_06032013.png]]
 +
<br.>
 +
<br.>
 +
[[Image: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===
 +
 
  
  

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June 3, 2013

Absorbance and Fluorescence Data

Finish preparing thiol DNA, AuNP and ThT sample<br.> 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.> 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