User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/07/12: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Meeting, DNA/AuNP Attachment</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Meeting, DNA/AuNP Attachment</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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*Centrifuge 4 times, extracting the supernatant each time and then redispersing in 250uL 50nM HEPES buffer | *Centrifuge 4 times, extracting the supernatant each time and then redispersing in 250uL 50nM HEPES buffer | ||
*Take absorbance of final solution and three supernatants | *Take absorbance of final solution and three supernatants | ||
==== | ====Notes==== | ||
Procedure stopped because the nanoparticles could not be extracted by centrifugation. Absorbance data to be taken tomorrow. | |||
===Procedure Update: Silica-coating AuNPs by MHA=== | ===Procedure Update: Silica-coating AuNPs by MHA=== | ||
We decided to follow the MHA procedure because MHA reduces the number of gold cores per silica-coated particles, and we would ideally have each individual gold nanoparticle covered by a layer of silica. Using the MHA versus not using it has also been very effective in the trials conducted so far | We decided to follow the MHA procedure because MHA reduces the number of gold cores per silica-coated particles, and we would ideally have each individual gold nanoparticle covered by a layer of silica. Using the MHA versus not using it has also been very effective in the trials conducted so far, and we think that using MHA will allow us to reach our desired optimal shell thickness of approximately 35nm.<br.> | ||
*Originally detailed on [http://openwetware.org/wiki/User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/05/24 05/24/2013] | *Originally detailed on [http://openwetware.org/wiki/User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/05/24 05/24/2013] | ||
*Day One: | *Day One: | ||
**Attach MHA to AuNPs: add 5mL of 10mM MHA to 5mL of AuNPs, stir overnight | **Attach MHA to AuNPs: add 5mL of 10mM MHA to 5mL of AuNPs, stir overnight | ||
***To prepare 10mM MHA solution: | |||
***(5 mL)×(L/1000mL)×(10mmoles/L)×(1mol/1000mmoles)×(288.49g/mol)=0.0144g MHA in 5mL 2-propanol | |||
***To prepare 5mL of 9.4nM AuNPs: | |||
***(29.67nM)V=(9.4nM)(5mL) | |||
***V=1.584mL AuNPs in 3.416mL PBS buffer | |||
***Combine MHA and AuNP solutions | |||
*Day Two: | *Day Two: | ||
**Prepare the following solutions: | **Prepare the following solutions: | ||
#1mL 9.4nM, 10nm AuNP&MHA/20μL TES/0.125mL NH4OH/5mL propanol | |||
#1mL 9.4nM, 10nm AuNP&MHA/15μL TES/0.125mL NH4OH/5mL propanol | |||
#1mL 9.4nM, 10nm AuNP&MHA/10μL TES/0.125mL NH4OH/5mL propanol | |||
#1mL 9.4nM, 10nm AuNP&MHA/5μL TES/0.125mL NH4OH/5mL propanol | #1mL 9.4nM, 10nm AuNP&MHA/5μL TES/0.125mL NH4OH/5mL propanol | ||
# | #*Stir solutions for one hour to allow reaction to proceed | ||
#*Centrifuge solutions at 4000 rpm for one hour | |||
#*Re-suspend particles in 140uL PBS (per solution) | |||
Latest revision as of 23:03, 26 September 2017
Meeting, DNA/AuNP Attachment | Main project page Previous entry Next entry |
July 12, 2013Notes from Meeting
DNA and AuNP Attachment ProcedureCalculations
NotesProcedure stopped because the nanoparticles could not be extracted by centrifugation. Absorbance data to be taken tomorrow. Procedure Update: Silica-coating AuNPs by MHAWe decided to follow the MHA procedure because MHA reduces the number of gold cores per silica-coated particles, and we would ideally have each individual gold nanoparticle covered by a layer of silica. Using the MHA versus not using it has also been very effective in the trials conducted so far, and we think that using MHA will allow us to reach our desired optimal shell thickness of approximately 35nm.<br.>
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