User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/04
|FCS Measurements and ZnPPIX Sample Prep||Main project page|
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June 4, 2013
Sonicate 10x diluted microspheres, dilute solutions 100x<br.> (1/10)V=(1/100)(30μL)<br.> V=3μL 10x diluted solution in 27μL distilled H2O
All samples prepared 06/03/2013, in addition to two modified microsphere solutions from Dr. Miller, were run on the FCS. Data was analyzed using N values, τ values, and C values were recorded. Tomorrow we will look in more detail at the time traces recorded directly from the instrument to look at differences in diffusion times.
Zinc Protoporphyrin Sample Prep: Calculations
Zinc protoporphyrin (ZnPPIX) samples will be prepared at the same concentrations, following the same procedure as the ThT samples.
(5μM)V=(4.44μM)(300μL)<br.> V=266.4μL 5μM ZnPPIX in 33.6μL<br.> (53.1μ)V=(2.2253μM)(300μL)<br.> V=12.57μL DNA in 287.43μL<br.>
2[ThT]:1[DNA]→ 2.068μM ZnPPIX<br.> 10% of 250μL=25μL (within 10% allows for accurate concentration comparison)<br.> Total solution volume: 275μL<br.> (2.068×10-6M)(275×10-6L)=5.31476×10-10moles ZnPPIX<br.> (5.31476×10-10moles)/(25×10-6L)=21.3μM ZnPPIX<br.> <br.> Make 21.3μM ZnPPIX:<br.> (5000μM)V=(21.3μM)(25μL)<br.> V=0.107μL in 24.9μL<br.> Add 25μL of 21.3μM ZnPPIX to 250μL DNA/AuNP solution<br.> <br.> Concentrations:<br.> DNA- 0.94μM<br.> AuNP- 12.55nM<br.> ZnPPIX- 1.9364μM<br.>
(5μM)V=(2.22μM)(300μL)<br.> V=133.2μL ZnPPIX in 166.8μL<br.>
Samples will be prepared and spectra taken tomorrow.
Modifying Silica Coated AuNPs with HS-DNA Procedure- Changed(?)