User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/05/29
DNA/ThT Reaction and Measurements | Main project page Previous entry Next entry |
May 29, 2013Reacting thiol DNA with ThTProcedurefrom Allison Alix 04/05/2013<br.>
new ThT concentration in solution:<br.> 5μM(200×10-6L)=0.001μmoles<br.> 0.001μmoles/(400×10-6L)=2.5μM<br.> <br.> thiol DNA concentration to 1.455 from 53.1:<br.> (53.1μM)V=(1.455μM)(200μL)<br.> V=5.48μL 53.1μM thiol DNA in 194μL filtered water(?)<br.> <br.> new thiol DNA concentration in solution:<br.> 1.455μM(200×10-6L)=2.91×10-4μmoles<br.> 2.91×10-4μmoles/(400×10-6L)=0.7275μM=727.5nM<br.> <br.>
DataAbsorbanceFlorescenceExcited at 414nm; peak observed ~500nm Reacting AuNPs with thiol DNA/ThT SolutionProcedurefrom Allison Alix 04/15/2013<br.> re-do of 05/28/2013<br.>
new DNA concentration: 727.5nM(250×10-6L)=0.181875nmoles<br.> 0.181875nmoles/(500×10-6L)=363.75nM<br.> new ThT concentration:<br.> 2.5μM(250×10-6L)=0.000625<br.> 0.000625/(500×10-6L)=1.25μM<br.>
DataAbsorbance<br.> Dark Blue: Water<br.> Light Blue: Thiol DNA, ThT, AuNP solution<br.> Red: Supernatant 1<br.> Green: Supernatant 2<br.> Purple: Supernatant 3<br.> ObservationsAgain, the extracted sample did not separate in the centrifuge as expected. I centrifuged as usual (though with larger time intervals and higher speeds) and removed the top "supernantant" layer despite the lack of separation. I took absorbance measurements, with inconclusive results. |