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

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{|{{table}} width="800"
{|{{table}} width="800"
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Silica Procedure Prep</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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NOTE: TES solution will be prepared immediately prior to use to minimize exposure
NOTE: TES solution will be prepared immediately prior to use to minimize exposure


'''Part 2: Preparing ZnPPIX solutions'''
'''Part 2: Preparing ZnPPIX solutions'''
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Zinc protoporphyrin (ZnPPIX) samples will be prepared at the same concentrations, following the same procedure as the ThT samples.
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
 
''2.22μM ZnPPIX and 1.11265μM DNA''
 


(5μM)V=(4.44μM)(300μL)
(5μM)V=(4.44μM)(300μL)
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V=12.57μL DNA in 287.43μL
V=12.57μL DNA in 287.43μL


ZnPPIX, DNA and AuNP
 
''ZnPPIX, DNA and AuNP''
 


2[ThT]:1[DNA]→ 2.068μM ZnPPIX
2[ThT]:1[DNA]→ 2.068μM ZnPPIX
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(5.31476×10-10moles)/(25×10-6L)=21.3μM ZnPPIX
(5.31476×10-10moles)/(25×10-6L)=21.3μM ZnPPIX


Make 21.3μM ZnPPIX:
* Make 21.3μM ZnPPIX:


(5000μM)V=(21.3μM)(25μL)
(5000μM)V=(21.3μM)(25μL)
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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:
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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
==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




Revision as of 11:40, 12 July 2013

Silica Procedure Prep <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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

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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