User:Allison K. Alix/Notebook/Thesis Research/2013/01/22

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Procedures

Hybridize Molecular Beacon (731nM) with the following @ same concentration in 1:1 ratios:

1) Elaine's Primer (307.8nM)

To get 307.8nM MB:

731nM(x) = (307.8nM)(5μL)

x = 2.105μL MB in 2.895μLH2O

mix with 5μL Elaines Primer and 180μL filtered H2O


2) Gold nano-particles (6.49nM)

To get 6.49nM MB:

731nm(x) = (6.49nM)(100μL)

x = 0.89μL MB in 99.11 μL H2O

mix with 100μL AuNP

_____

  • Add 500μL H2O to 4 repeating and 8 repeating telomeric DNA strands to get the following concentrations:

1) 4 telomeric repeats:

132.8nmoles/0.5mL x 1000ml/1L = 265.6μM

2) 8 telomeric repeats:

71.4nmoles/0.5mL x 1000mL/1L = 142.8μM

  • Hybridize ZnPPIX (5μM) with each DNA strand:

1) 4 telomeric repeats:

To get 5μM :

265.6 μM (x) = 5μM (100μL)

x = 1.88 μL in 98.12μL H2O

mix with 100μL ZnPPIX

2) 8 telomeric repeats:

To get 5μM :

142.8 μM (x) = 5μM (100μL)

x = 3.5 μL in 96.5 μL H2O

mix with 100μL ZnPPIX

Take Absorbance/Fluorescence of all 4 hybridizations