User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/07/22: Difference between revisions

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==July 22, 2013==
==July 22, 2013==
===Procedure Update: Silica-coating AuNPs by MHA, Round 2===
===Procedure: Silica-coating AuNPs by MHA, Day One Round 2===
*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:
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***Combine MHA and AuNP solutions
***Combine MHA and AuNP solutions
**Note: two samples made, one made in isopropanol and the other in PBS
**Note: two samples made, one made in isopropanol and the other in PBS
*Day Two:
**Prepare the following solutions:
#1mL 9.4nM, 9.65nm AuNP&MHA, isopropanol/4μL TES/0.125mL NH4OH/5mL propanol
#1mL 9.4nM, 9.65nm AuNP&MHA, isopropanol/3μL TES/0.125mL NH4OH/5mL propanol
#1mL 9.4nM, 9.65nm AuNP&MHA, isopropanol/2μL TES/0.125mL NH4OH/5mL propanol
#1mL 9.4nM, 9.65nm AuNP&MHA, isopropanol/1μL TES/0.125mL NH4OH/5mL propanol
#1mL 9.4nM, 9.65nm AuNP&MHA, PBS/4μL TES/0.125mL NH4OH/5mL propanol
#1mL 9.4nM, 9.65nm AuNP&MHA, PBS/3μL TES/0.125mL NH4OH/5mL propanol
#1mL 9.4nM, 9.65nm AuNP&MHA, PBS/2μL TES/0.125mL NH4OH/5mL propanol
#1mL 9.4nM, 9.65nm AuNP&MHA, PBS/1μL 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)
#**Dilute each solution 10 and 100 times




Revision as of 14:42, 22 July 2013

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July 22, 2013

Procedure: Silica-coating AuNPs by MHA, Day One Round 2

  • Originally detailed on 05/24/2013
  • Day One:
    • 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:
      • (23.5nM)V=(9.4nM)(5mL)
      • V=2mL 23.5nM 9.65nm AuNPs in 3mL PBS buffer
      • Combine MHA and AuNP solutions
    • Note: two samples made, one made in isopropanol and the other in PBS



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