User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/04/02

From OpenWetWare

< User:Madeleine Y. Bee | Notebook | Single Molecule Fluorescence | 2013 | 04(Difference between revisions)
Jump to: navigation, search
(Procedures)
Current revision (16:21, 12 July 2013) (view source)
(April 2, 2013)
 
(3 intermediate revisions not shown.)
Line 7: Line 7:
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
==April 2, 2013==
==April 2, 2013==
 +
=====Hybridize thiol DNA and Molecular Beacon, then AuNP=====
 +
DNA: 53.1μM → 727.5nM
 +
<br.>
 +
(53.1μM)V=(500μL)(1455nM)<br.>
 +
V=13.7μL 53.1μM DNA in 486.3μL filtered H<sub>2</sub>O <br.>
 +
(1455nM)V=(200μL)(727.5nM)<br.>
 +
V=100μL 1455nM DNA in 100μL H<sub>2</sub>O <br.>
 +
MB: 7.31μM → 800nM<br.>
 +
 +
(7130nM)V=(800nM)(50μL)<br.>
 +
V=5.61μL 7.31μM MB in 44.53μL H<sub>2</sub>O<br.>
 +
<br.>
 +
Combine 50μL 727.5nM thiol DNA with 50μL 800nM MB, heat at 75°C for 25 minutes, cool to room temperature.<br.>
 +
 +
=====Hybridize thiol DNA and ThT, then AuNP=====
 +
ThT: from January 29, 2013- assumed 100%, Dr. Miller's 5mM ThT used in this procedure<br.>
 +
<br.>
 +
(5mM)V=(5μM)(10mL)<br.>
 +
V= 10μL of 5mM ThT in 9990μL PBS buffer<br.>
 +
(5μM)V=(800nM)(1mL)<br.>
 +
V= 160μL 5μM ThT in 840μL PBS buffer<br.>
 +
<br.>
 +
Combine 50μL 727.5nM thiol DNA with 50μL 800nM ThT<br.>
 +
 +
=====AuNP with Ethanethiol=====
 +
AuNP radius: (from Dr. Miller March 1, 2013) 14nm diameter=7nm radius<br.>
 +
Area of AuNP: 4πr<sup>2</sup>=615.75 nm<sup>2</sup><br.>
 +
Sulfur atom radius: 200pm diameter=100pm radius<br.>
 +
Area of sulfur atom: r<sup>2</sup>=40000pm<sup>2</sup><br.>
 +
sulfur atoms/AuNP=(6.1575×10<sup>-16</sup>m<sup>2</sup>)/(4×10<sup>-20</sup>m<sup>2</sup>)=15393<br.>
 +
<br.>
 +
AuNP: 19.4nM<br.>
 +
Ethanethiol: make 298μM, density = 0.839g/mL<br.>
 +
<br.>
 +
2.5μL × (10<sup>-6</sup>L/1μL) × (1000mL/1L) × (0.839g/1mL) × (1 mol/ 62.134g) = 3.376×10<sup>-4</sup>mol = 0.34 M = 340 mM<br.>
 +
<br.>
 +
(1mM)V=(340mM)(1mL)<br.>

Current revision

Project name Main project page
Previous entry      Next entry

April 2, 2013

Hybridize thiol DNA and Molecular Beacon, then AuNP

DNA: 53.1μM → 727.5nM
(53.1μM)V=(500μL)(1455nM)
V=13.7μL 53.1μM DNA in 486.3μL filtered H2O
(1455nM)V=(200μL)(727.5nM)
V=100μL 1455nM DNA in 100μL H2O

MB: 7.31μM → 800nM

(7130nM)V=(800nM)(50μL)
V=5.61μL 7.31μM MB in 44.53μL H2O

Combine 50μL 727.5nM thiol DNA with 50μL 800nM MB, heat at 75°C for 25 minutes, cool to room temperature.

Hybridize thiol DNA and ThT, then AuNP

ThT: from January 29, 2013- assumed 100%, Dr. Miller's 5mM ThT used in this procedure

(5mM)V=(5μM)(10mL)
V= 10μL of 5mM ThT in 9990μL PBS buffer
(5μM)V=(800nM)(1mL)
V= 160μL 5μM ThT in 840μL PBS buffer

Combine 50μL 727.5nM thiol DNA with 50μL 800nM ThT

AuNP with Ethanethiol

AuNP radius: (from Dr. Miller March 1, 2013) 14nm diameter=7nm radius
Area of AuNP: 4πr2=615.75 nm2
Sulfur atom radius: 200pm diameter=100pm radius
Area of sulfur atom: r2=40000pm2
sulfur atoms/AuNP=(6.1575×10-16m2)/(4×10-20m2)=15393

AuNP: 19.4nM
Ethanethiol: make 298μM, density = 0.839g/mL

2.5μL × (10-6L/1μL) × (1000mL/1L) × (0.839g/1mL) × (1 mol/ 62.134g) = 3.376×10-4mol = 0.34 M = 340 mM

(1mM)V=(340mM)(1mL)



Personal tools