User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/03/19: Difference between revisions
From OpenWetWare
(4 intermediate revisions by the same user 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. ##### --> | ||
==March 19, 2013== | ==March 19, 2013== | ||
====Hybridization of thiol DNA, AuNP, and Molecular Beacon==== | |||
DNA:AuNP ratio 75:1<br.> | |||
AuNP concentration: 19.4nM<br.> | |||
thiol DNA concentration: 1455nM<br.> | |||
#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<br.> | |||
#Molecular Beacon concentration: 7.31μM = 7310nM | |||
(7310)V=(800nM)(150μL)<br.> | |||
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<br.> | |||
heated for approximately 25 minutes at 70°C; cooled to room temperature<br.> | |||
=====Procedure===== | |||
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
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-
|