User:Allison K. Alix/Notebook/Thesis Research/2013/08/27

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 Revision as of 11:41, 27 August 2013 (view source) (→Procedures)← Previous diff Current revision (10:54, 28 August 2013) (view source) (→Abs/Fluorescence of Au@SiO2/DNA-ThT Sample) (6 intermediate revisions not shown.) Line 31: Line 31: [ThT] must be twice what [DNA] is [ThT] must be twice what [DNA] is - [DNA] = 1.11225μM, so [ThT] must be 2.2245μM + [DNA] = 0.624μM, so [ThT] must be 1.248μM - Use 250μL of DNA-AuNP solution (10%=25μM) + Use 150μL of DNA-AuNP solution (10%=15μL) - 2.2245x10-6M x 275x10-6L solution = 6.117375x10-10 moles ThT + 1.248x10-6M x 165x10-6L solution = 2.0592x10-10 moles ThT - (6.117375x10-10 moles ThT)/ 25 x 10-6 L = 24.4695μM + (2.0592x10-10 moles ThT)/ 15 x 10-6 L = 13.728μM - -Add 25μL of 24.4695μM ThT to 250μL thiol-DNA/AuNP solution + -Add 15μL of 13.728μM to 150μL thiol-DNA/AuNP solution NEW Concentrations: NEW Concentrations: - [DNA]= 1.011μM [AuNP] = 13.486μM [ThT] = 2.2245μM + [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. + +

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To Do

• Align using YG Fluorescent Beads
• Add ThT to Au@SiO2/DNA solution
• Obtain UV-Vis and Fluorescence of above solution

Au@SiO2/DNA UV-Vis Data

ε of thiol-DNA= 607500 A=0.379 b=1 cm

A=εbc

c=A/εb

c = 6.23x10^-7M = 0.624uM

[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.