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

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(Notes)
(Notes)
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* Dilute beads 10X and 100X more, there was WAY too much signal and no curve seen in FCS measurements
* Dilute beads 10X and 100X more, there was WAY too much signal and no curve seen in FCS measurements
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Revision as of 13:34, 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

Data

Image:Pic_of_ZnPPIX_abs.png

Image:Pic_of_ZnPPIX_fluor.png

Notes

To do:

  • Prepare new ZnPPIX/DNA sample and take fluorescence (it is curious that there was no fluorescence seen in this sample)
  • Prepare AuNP/ZnPPIX sample and take fluorescence (be sure that the fluorescence seen in the AuNP/DNA/ZnPPIX sample is not due to an AuNP/ZnPPIX interaction)
  • Dilute beads 10X and 100X more, there was WAY too much signal and no curve seen in FCS measurements




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