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

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{|{{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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2.22μM ZnPPIX and 1.11265μM DNA
''2.22μM ZnPPIX and 1.11265μM DNA''




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




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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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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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