User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/03/19: Difference between revisions
From OpenWetWare
| Line 8: | Line 8: | ||
==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===== | =====Procedure===== | ||
thiol DNA and AuNP solution- | thiol DNA and AuNP solution- | ||
Revision as of 13:17, 12 July 2013
 Project name
|
<html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
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- 1. 500μL 2910nM DNA and 500μL 4% TEA with 100nM DTT; allowed to react for 10 minutes 2. 16mL (in 4mL aliquots) ethyl acetate used to extract (done under the fume hood) 3. 464.8μL 727.5nM DNA combined with 464.8μL 9.7nM AuNP and 70.4μL sodium citrate buffer; allowed to react for 10 minutes 4. centrifuge for 15-25 minutes
| |
