User:Allison K. Alix/Notebook/Thesis Research/2013/05/29
From OpenWetWare
DNA/AuNP/MB Preparation | Main project page Previous entry Next entry |
Objectives
ProceduresPart 1: Preparation of 100mM DTT in 4% TEA solution Original DTT concentration 1M 1M(x) = (100x10-3M)(1mL) x = 0.1mL = 100μL in 900μL 4% TEA 4%(900μL) = 36μL TEA in 864μL PBS OVERALL: 100μL 1M DTT + 36μL TEA + 864μL PBS Part 2: Prepare DNA-AuNP to bind MB to 1) Mix 100μL 2.91μM thiol-DNA with 100μL 100mM DTT in 4% TEA. React 10 minutes [DNA]=1.45μM 2) Extract DTT using 4 2mL aliquots of ethyl acetate 3) Mix 93μL thiol-DNA with 93μL 19.4nM AuNP and 14μL sodium citrate buffer. React 10 minutes. [DNA] = 727.5nM [AuNP] = 9.7nM 4) Centrifuge and redisperse in 50mM HEPES buffer 5) Mix 20μL of 7.61μM MB with 100μL of thiol DNA/AuNP mixture 6) Heat @75°C for 25 minutes. 7) Observe absorbance and fluorescence spectra
|