User:Allison K. Alix/Notebook/Thesis Research/2013/08/29: Difference between revisions

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==New Procedure==
==New Procedure==


1) Combine 1mL 5nM silica coated Au particles with 5uL 1mM APS. Stir for 1 hour.
1) Combine 1mL 5nM silica coated Au particles with 5uL APS. Stir for 1 hour.
 
*Concentration of APS not noted. any excess APS will be removed after sentrifugation


2) Add 2mg Sulfo-SMCC in 3mL KPS buffer (pH =7.4, 0.05M) add 1.5mL to NH2 functionalized AuNP's.
2) Add 2mg Sulfo-SMCC in 3mL KPS buffer (pH =7.4, 0.05M) add 1.5mL to NH2 functionalized AuNP's.


3) Stir for 1hr. and centrifuge for ??
3) Stir for 1hr. and centrifuge for 30 seconds at a time at 9000rpm for three times. Remove supernatant and redisperse in 500uL KP buffer. Also centrifuge superntants and redisperse in ~30uL KP buffer. New volume of solution: 530uL


4) Add  
4) Add 270uL 1.5uM HS-DNA. Stir for 2 hours.




Revision as of 10:59, 29 August 2013

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Revisions to DNA attachment on Au@SiO2

  • KP buffer was NOT at 7.4 (was at 7.13). pH was readjusted to 7.4 using ~2M NaOH.
  • One step of the procedure notes to centrifuge DNA and Sulfo-SMCC to remove excess SSMCC; however, after centrifugation, no separation between the two was observed. Because of this, it will be better to add the Sulfo-SMCC to the NH2 functionalized Au@SiO2 and then add the DNA last.
  • Procedure 1 has too many rinsing steps which lead me to believe many of the nanoparticles are lost.

New Procedure

1) Combine 1mL 5nM silica coated Au particles with 5uL APS. Stir for 1 hour.

  • Concentration of APS not noted. any excess APS will be removed after sentrifugation

2) Add 2mg Sulfo-SMCC in 3mL KPS buffer (pH =7.4, 0.05M) add 1.5mL to NH2 functionalized AuNP's.

3) Stir for 1hr. and centrifuge for 30 seconds at a time at 9000rpm for three times. Remove supernatant and redisperse in 500uL KP buffer. Also centrifuge superntants and redisperse in ~30uL KP buffer. New volume of solution: 530uL

4) Add 270uL 1.5uM HS-DNA. Stir for 2 hours.



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