User:Allison K. Alix/Notebook/Thesis Research/2013/06/05

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(Procedures)
(Procedures)
Line 28: Line 28:
(5μM)V=(4.44μM)(300μL)
(5μM)V=(4.44μM)(300μL)
 +
V=266.4μL 5μM ZnPPIX in 33.6μL
V=266.4μL 5μM ZnPPIX in 33.6μL
 +
(53.1μ)V=(2.2253μM)(300μL)
(53.1μ)V=(2.2253μM)(300μL)
 +
V=12.57μL DNA in 287.43μL
V=12.57μL DNA in 287.43μL
Line 35: Line 38:
2[ThT]:1[DNA]→ 2.068μM ZnPPIX
2[ThT]:1[DNA]→ 2.068μM ZnPPIX
 +
10% of 250μL=25μL (within 10% allows for accurate concentration comparison)
10% of 250μL=25μL (within 10% allows for accurate concentration comparison)
 +
Total solution volume: 275μL
Total solution volume: 275μL
 +
(2.068×10-6M)(275×10-6L)=5.31476×10-10moles ZnPPIX
(2.068×10-6M)(275×10-6L)=5.31476×10-10moles ZnPPIX
 +
(5.31476×10-10moles)/(25×10-6L)=21.3μM ZnPPIX
(5.31476×10-10moles)/(25×10-6L)=21.3μM ZnPPIX
Line 43: Line 50:
(5000μM)V=(21.3μM)(25μL)
(5000μM)V=(21.3μM)(25μL)
 +
V=0.107μL in 24.9μL
V=0.107μL in 24.9μL
 +
Add 25μL of 21.3μM ZnPPIX to 250μL DNA/AuNP solution
Add 25μL of 21.3μM ZnPPIX to 250μL DNA/AuNP solution
Concentrations:
Concentrations:
 +
DNA- 0.94μM
DNA- 0.94μM
 +
AuNP- 12.55nM
AuNP- 12.55nM
 +
ZnPPIX- 1.9364μM
ZnPPIX- 1.9364μM
 +
2.22μM ZnPPIX
2.22μM ZnPPIX
 +
(5μM)V=(2.22μM)(300μL)
(5μM)V=(2.22μM)(300μL)
 +
V=133.2μL ZnPPIX in 166.8μL
V=133.2μL ZnPPIX in 166.8μL

Revision as of 11:51, 5 June 2013

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Objectives

Procedures

Part 1: Preparing solutions for coating AuNP with Silica

1) dissolve 0.716g PVP (10kg/mol) in 10mL water (0.00716M)

2) Dissolve 0.42mL NH4OH (29.3wt% NH3 in H2O) in 9.58mL Ethanol (200 proof) (4.2vol%)

NOTE: TES solution will be prepared immediately prior to use to minimize exposure

Part 2: Preparing ZnPPIX solutions

Zinc protoporphyrin (ZnPPIX) samples will be prepared at the same concentrations, following the same procedure as the ThT samples.

2.22μM ZnPPIX and 1.11265μM DNA

(5μM)V=(4.44μM)(300μL)

V=266.4μL 5μM ZnPPIX in 33.6μL

(53.1μ)V=(2.2253μM)(300μL)

V=12.57μL DNA in 287.43μL

ZnPPIX, DNA and AuNP

2[ThT]:1[DNA]→ 2.068μM ZnPPIX

10% of 250μL=25μL (within 10% allows for accurate concentration comparison)

Total solution volume: 275μL

(2.068×10-6M)(275×10-6L)=5.31476×10-10moles ZnPPIX

(5.31476×10-10moles)/(25×10-6L)=21.3μM ZnPPIX

Make 21.3μM ZnPPIX:

(5000μM)V=(21.3μM)(25μL)

V=0.107μL in 24.9μL

Add 25μL of 21.3μM ZnPPIX to 250μL DNA/AuNP solution

Concentrations:

DNA- 0.94μM

AuNP- 12.55nM

ZnPPIX- 1.9364μM

2.22μM ZnPPIX

(5μM)V=(2.22μM)(300μL)

V=133.2μL ZnPPIX in 166.8μL



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