User:Allison K. Alix/Notebook/Thesis Research/2013/01/22: Difference between revisions
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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 2.895μLH<sub>2</sub>O | |||
mix with 5μL Elaines Primer and 180μL filtered H<sub>2</sub>O | |||
2) Gold nano-particles (6.49nM) | |||
To get 6.49nM MB: | |||
731nm(x) = (6.49nM)(100μL) | |||
x = 0.89μL MB in 99.11 μL H<sub>2</sub>O | |||
mix with 100μL AuNP | |||
_____ | |||
* Add 500μL H<sub>2</sub>O 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 H<sub>2</sub>O | |||
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 H<sub>2</sub>O | |||
mix with 100μL ZnPPIX | |||
Take Absorbance/Fluorescence of all 4 hybridizations | |||
Revision as of 11:43, 22 January 2013
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ProceduresHybridize 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
To get 6.49nM MB: 731nm(x) = (6.49nM)(100μL) x = 0.89μL MB in 99.11 μL H2O mix with 100μL AuNP _____
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
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
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