User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/04/02: Difference between revisions
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==April 2, 2013== | ==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 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 | |||
=====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 | |||
Revision as of 08:07, 23 April 2013
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April 2, 2013Hybridize thiol DNA and Molecular Beacon, then AuNPDNA: 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 AuNPThT: 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 EthanethiolAuNP radius: (from Dr. Miller March 1, 2013) 14nm diameter=7nm radius
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