User:Allison K. Alix/Notebook/Thesis Research/2013/05/23

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Observe Fluor w/ and w/o AuNP Main project page
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Objectives

  • Take UV-Vis/Fluorescence Measurements of same DNA/ThT concentration in DNA-AuNP-ThT mixture
  • Observe where fluorescence quenching is occuring

Procedures

Part 1: Adding ThT to thiol-DNA/AuNP

[ThT] must be twice what [DNA] is

[DNA] = 1.11225μM, so [ThT] must be 2.2245μM

Use 250μL of DNA-AuNP solution (10%=25μM)

2.2245x10-6M x 275x10-6L solution = 6.117375x10-10 moles ThT

(6.117375x10-10 moles ThT)/ 25 x 10-6 L = 24.4695μM

-Add 25μL of 24.4695μM ThT to 250μL thiol-DNA/AuNP solution

NEW Concentrations:

[DNA]= 1.011μM [AuNP] = 13.486μM [ThT] = 2.2245μM

Take Fluorescence Spectrum

Part 2: Preparing thiol-DNA/ThT solution using same concentrations in thiol-DNA/AuNP/ThT solution

1) Prepare 250μL of 2.022μM thiol DNA

(5.82μM)(x) = (250μL)(2.022μM)

x = 86.85 μL 5.82μM thiol-DNA in 163.15μL PBS Buffer

2) Prepare 250μL of 4.449μM ThT

(5μM)(x) = (4.449μM)(250μL)

x = 222.45μL in 27.55μL PBS Buffer

3) Mix 250μL 2.002μM thiol-DNA with 250μL 4.449μM ThT

4) Obtain Fluorescence Spectrum

Data

Image:Fluor_pic05232013.png




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