User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/03/19: Difference between revisions
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==March 19, 2013== | ==March 19, 2013== | ||
====Hybridization of thiol DNA, AuNP, and Molecular Beacon==== | ====Hybridization of thiol DNA, AuNP, and Molecular Beacon==== | ||
DNA:AuNP ratio 75:1 | DNA:AuNP ratio 75:1<br.> | ||
AuNP concentration: 19.4nM | AuNP concentration: 19.4nM<br.> | ||
thiol DNA concentration: 1455nM | thiol DNA concentration: 1455nM<br.> | ||
#100μL 5.5μM ThT, 100μL 727.5nM thiol DNA, and 9.7nM AuNP | #100μL 5.5μM ThT, 100μL 727.5nM thiol DNA, and 9.7nM AuNP | ||
heated for approximately 25 minutes at 70°C; cooled to room temperature | heated for approximately 25 minutes at 70°C; cooled to room temperature<br.> | ||
#Molecular Beacon concentration: 7.31μM = 7310nM | #Molecular Beacon concentration: 7.31μM = 7310nM | ||
(7310)V=(800nM)(150μL) | (7310)V=(800nM)(150μL)<br.> | ||
V= 5.5μL MB in 44.5μL filtered H<sub>2</sub>O | V= 5.5μL MB in 44.5μL filtered H<sub>2</sub>O<br.> | ||
Combine: 50μL 800nM MB, 50μL 727.5nM thiol DNA with 9.7nM AuNP | Combine: 50μL 800nM MB, 50μL 727.5nM thiol DNA with 9.7nM AuNP<br.> | ||
heated for approximately 25 minutes at 70°C; cooled to room temperature | heated for approximately 25 minutes at 70°C; cooled to room temperature<br.> | ||
=====Procedure===== | |||
thiol DNA and AuNP solution- | thiol DNA and AuNP solution- | ||
#500μL 2910nM DNA and 500μL 4% TEA with 100nM DTT; allowed to react for 10 minutes | |||
#16mL (in 4mL aliquots) ethyl acetate used to extract (done under the fume hood) | |||
#464.8μL 727.5nM DNA combined with 464.8μL 9.7nM AuNP and 70.4μL sodium citrate buffer; allowed to react for 10 minutes | |||
#Centrifuge for 15-25 minutes | |||
Revision as of 13:18, 12 July 2013
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March 19, 2013Hybridization of thiol DNA, AuNP, and Molecular BeaconDNA:AuNP ratio 75:1<br.> AuNP concentration: 19.4nM<br.> thiol DNA concentration: 1455nM<br.>
heated for approximately 25 minutes at 70°C; cooled to room temperature<br.>
(7310)V=(800nM)(150μL)<br.> V= 5.5μL MB in 44.5μL filtered H2O<br.> Combine: 50μL 800nM MB, 50μL 727.5nM thiol DNA with 9.7nM AuNP<br.> heated for approximately 25 minutes at 70°C; cooled to room temperature<br.> Procedurethiol DNA and AuNP solution-
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