User:Allison K. Alix/Notebook/Thesis Research/2013/05/31: Difference between revisions
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8) Centrifuge for 7 minutes at 10000rpm | 8) Centrifuge for 7 minutes at 10000rpm | ||
Part 2: Preparation of | Part 2: Preparation of thiol-DNA/ThT solution | ||
concentration of DNA must match [DNA] in AuNP/DNA solution and [ThT] must be at least twice this | |||
1) 5.82μM(x)=(2.2253μM)(250μL) | |||
x = 95.6μL in 154.4μL PBS to make 250μL 2.2263μM DNA | |||
2) 120μM(x)=(250μL)(4.44μM) | |||
x = 9.25μL in 240.75μL PBS to make 250μL 4.44μM ThT | |||
Revision as of 12:19, 3 June 2013
 Project name
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Objectives
ProceduresPart 1: Preparation of AuNP-DNA sample Final concentrations of AuNP and DNA should be 14.835nM and 1.11625μM respectively 1) 53.1μM(x)=(4.4506μM)(250μL) x= 20.95μL in 229.05μL PBS mix 20.95μL 53.1μM thiol-DNa with 229.05μL PBS 2) Mix 250μL DNA solution with 250μL 100mM DTT in 4% TEA NEW [DNA] = 2.225μM 3) React 10 minutes 5) Extract DTT from solution using 4 2mL aliquots of ethyl acetate 6) Mix 232.4μL DNA, 232.4μL 29.7nM AuNP, and 36.2μL sodium citrate buffer NEW [DNA] = 1.034μM NEW {[AuNP] = 13.805nM 7) React 10 minutes 8) Centrifuge for 7 minutes at 10000rpm Part 2: Preparation of thiol-DNA/ThT solution concentration of DNA must match [DNA] in AuNP/DNA solution and [ThT] must be at least twice this 1) 5.82μM(x)=(2.2253μM)(250μL) x = 95.6μL in 154.4μL PBS to make 250μL 2.2263μM DNA 2) 120μM(x)=(250μL)(4.44μM) x = 9.25μL in 240.75μL PBS to make 250μL 4.44μM ThT
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