User:Allison K. Alix/Notebook/Thesis Research/2013/05/14: Difference between revisions
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5) 5μL supernatant 3 with 1μL loading buffer | 5) 5μL supernatant 3 with 1μL loading buffer | ||
==Data== | |||
Revision as of 10:15, 14 May 2013
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Objectives
Procedures
Previous concentrations used:
To be used:
Calculations: Preparing 17.8μM thiol-DNA original concentration of thiol-DNA- 53.1μM 53.1μM (x) = (17.8μM)(250μL) x= 83.8μL in 166.2μL buffer Preparing 20μM ThT original conc ThT= 5mM 5mM (x) = (0.02mM)(10mL) x=40μL in 9960μL H2O
1) Mix 250μL 17.8μM thiol-DNA with 250μL 20μM ThT
2) Heat at 75°C for ~25min 3) Cool to room temperature 4) Take UV-Vis/Fluorescence measurements of thiol-DNA/ThT sample Part 2: Hybridizing thiol-DNA/ThT with AuNP 1) Mix 250 μL 8.9μM thiol-DNA/10μM ThT solution with 250μL 4%TEA containing 100mM DTT
2) Extract DTT using 4 2mL aliquots of ethyl acetate 3) Mix 232.4μL of thiol-DNA/ThT with 232.4μL 29.67nM AuNP and 35.2μL sodium citrate buffer
Part 3: Gel Electrophoresis (2% Agarose) Prepare the following: 1) 5μL 100bp ladder with 1μL loading buffer 2) 5μL thiol-DNA/ThT/AuNP with 1μL loading buffer 3) 5μL supernatant 1 with 1μL loading buffer 4) 5μL supernatant 2 with 1μL loading buffer 5) 5μL supernatant 3 with 1μL loading buffer Data |