User:Allison K. Alix/Notebook/Thesis Research/2013/08/27: Difference between revisions
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[[Image:DNA_method_2.jpg]] | [[Image:DNA_method_2.jpg]] | ||
ε of thiol-DNA= 607500 | |||
A=0.379 | |||
b=1 cm | |||
A=εbc | |||
c=A/εb | |||
c = 6.23x10^-7M = 0.624uM | |||
==Addition of ThT== | |||
[ThT] must be twice what [DNA] is | |||
[DNA] = 0.624μM, so [ThT] must be 1.248μM | |||
Use 150μL of DNA-AuNP solution (10%=15μL) | |||
1.248x10-6M x 165x10-6L solution = 2.0592x10-10 moles ThT | |||
(2.0592x10-10 moles ThT)/ 15 x 10-6 L = 13.728μM | |||
-Add 15μL of 13.728μM to 150μL thiol-DNA/AuNP solution | |||
NEW Concentrations: | |||
[DNA]= 0.567μM, [ThT] = 1.248μM | |||
1) Prepare 100uL 13.728uM ThT from 20uM ThT (prepared 5/14/13) | |||
13.728uM(100uL) = 20uM(x) | |||
x = 68.64uL in 31.36uL PBS | |||
==Abs/Fluorescence of Au@SiO2/DNA-ThT Sample== | |||
[[Image:AuSiO2DNAThT_OWW.png]] | |||
[[Image:AuSiO2DNAThTFluorOWW.png]] | |||
Intensity is far lower than expected for fluorescence. It is possible that silica coating is not preventing quenching and we are in need of a thicker coating. Will allow silica to stir for longer than previously stirred and possibly consider a higher volume of TES. | |||
Revision as of 07:54, 28 August 2013
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To Do
Au@SiO2/DNA UV-Vis Dataε of thiol-DNA= 607500 A=0.379 b=1 cm A=εbc c=A/εb c = 6.23x10^-7M = 0.624uM Addition of ThT[ThT] must be twice what [DNA] is [DNA] = 0.624μM, so [ThT] must be 1.248μM Use 150μL of DNA-AuNP solution (10%=15μL) 1.248x10-6M x 165x10-6L solution = 2.0592x10-10 moles ThT (2.0592x10-10 moles ThT)/ 15 x 10-6 L = 13.728μM -Add 15μL of 13.728μM to 150μL thiol-DNA/AuNP solution NEW Concentrations: [DNA]= 0.567μM, [ThT] = 1.248μM 1) Prepare 100uL 13.728uM ThT from 20uM ThT (prepared 5/14/13) 13.728uM(100uL) = 20uM(x) x = 68.64uL in 31.36uL PBS Abs/Fluorescence of Au@SiO2/DNA-ThT Sample
Intensity is far lower than expected for fluorescence. It is possible that silica coating is not preventing quenching and we are in need of a thicker coating. Will allow silica to stir for longer than previously stirred and possibly consider a higher volume of TES.
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