Difference between revisions of "User:Allison K. Alix/Notebook/Thesis Research/2013/08/12"

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(AuNP@SiO2/DNA attachment)
m (AuNP@SiO2/DNA attachment)
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==AuNP@SiO2/DNA attachment==
==AuNP@SiO2/DNA attachment==
adapted from Madeleine Y. Bee[[http://openwetware.org/wiki/User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/06/10]]
adapted from Madeleine Y. Bee[http://openwetware.org/wiki/User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/06/10]
1) Prepare a fresh 10X dilution of AuNP@SiO2
1) Prepare a fresh 10X dilution of AuNP@SiO2

Revision as of 07:28, 12 August 2013

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To Do

  • AuNP@SiO2/DNA attachment procedure (10X diluted AuNP@SiO2 prepared with 2uL TES to be used)

AuNP@SiO2/DNA attachment

adapted from Madeleine Y. Bee[1]

1) Prepare a fresh 10X dilution of AuNP@SiO2

  • Sonicate AuNP@SiO2 for 10 minutes
  • Mix 15uL AuNP@SiO2 with 135uL filter milliQ water (@ room temperature), vortex to mix thorooughly

2) Mix 150uL AuNP@SiO2 with 15uL APS and 235uL 100% ethanol. Sonicate for 30 minutes

3) Centrifuge solution a total of 5 times at 9100rpm for 5 minutes. Redisperse in ethanol for the first three and 10mM sodium phosphate buffer (pH=9) for the last two.

NOTE: after first round of centrifugation, it was apparent 5 minutes was a sufficient amount of time. However, after removal of the supernatant and redispersion in ethanol, there was difficulty getting the particles back into solution. They were allowed to sonicate for 10 minutes to try to induce separation.