User:Allison K. Alix/Notebook/Thesis Research/2013/07/15

From OpenWetWare
Jump to navigationJump to search
Owwnotebook icon.png DNA/AuNP Attachment & Silica Coating Pt 1 Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

To Do

  • Use both 268mM sodium citrate buffer and 100mM citrate buffer in AuNP/DNA attachment procedures
  • Take UV-Vis spectra of each sample
  • Determine concentration of DNA in solution with AuNP as well as in supernantants
  • Begin MHA Silica Coating Procedure

DNA/AuNP Attachment

1) Prepare 2 solutions containing:

  • 250uL 4.45uM thiol DNA + 250uL 100mM DTT in 4% TEA

2) React 10 minutes and extract DNA using ethyl acetate

3) Prepare the following two solutions

A) 240.7uL 29.67nM AuNP + 240.7uL 2.225uM DNA + 18.7uL sodium citrate

  • Final concentrations: [AuNP]=14.28nM, [DNA]=1.071uM, [sodium citrate]=10mM

B) 157.5uL 29.67nM auNP + 157.5uL 2.225uM DNA + 35uL sodium citrate

  • Final Concentrations: [AuNP]= 13.35nM, [DNA]=1.0014uM, [sodium citrate]=10mM

4) React 10 minutes

5) Centrifuge for 4 minutes at 7000rpm, redisperse in 50mM HEPES buffer

Silica Coating Procedure: Part 1

  • We have decided to use the procedure adapted from [1] to coat the AuNP with Silica. The first step is to attach a ligand stabilizer, mercaptohexadecanoic acid, MHA, to the nanoparticles, as described below.

1) Dissolve 0.0144g MHA in 5mL PBS (10mM MHA), combine with 5mL AuNPs

2) Stir overnight

  • Part 2 (addition of TEOS in propanol/ammonia in water will be completed tomorrow)