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

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(Procedures)
(April 2, 2013)
Line 7: Line 7:
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==April 2, 2013==
==April 2, 2013==
 +
=====Hybridize thiol DNA and Molecular Beacon, then AuNP=====
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DNA: 53.1μM → 727.5nM
 +
 +
(53.1μM)V=(500μL)(1455nM)
 +
V=13.7μL 53.1μM DNA in 486.3μL filtered H<sub>2</sub>O
 +
(1455nM)V=(200μL)(727.5nM)
 +
V=100μL 1455nM DNA in 100μL H<sub>2</sub>O
 +
 +
MB: 7.31μM → 800nM
 +
 +
(7130nM)V=(800nM)(50μL)
 +
V=5.61μL 7.31μM MB in 44.53μL H<sub>2</sub>O
 +
 +
Combine 50μL 727.5nM thiol DNA with 50μL 800nM MB, heat at 75°C for 25 minutes, cool to room temperature
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 +
=====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=====
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AuNP radius: (from Dr. Miller March 1, 2013) 14nm diameter=7nm radius

Revision as of 10:07, 23 April 2013

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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



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