User:Allison K. Alix/Notebook/Thesis Research/2013/07/10

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DNA-AuNP attachment

  • This will be used to calculate the amount of DNA that is adsorbing onto the gold nanoparticles. I will be able to compare this amount to the amount being adsorbed onto the silica coated gold nanoparticles to get an idea of whether or not the silica hinders that attachment and if so by how much. Will I need to add more DNA?

Follow procedure from User:Allison_K._Alix/Notebook/Thesis_Research/2013/04/15

Procedures:

1) Mix 250uL 4.405uM DNA with 250uL 100mM DTT in 4% TEA

2) React for 10 minutes

3) Extract DNA using 4 2mL aliquots of ethyl acetate

4) Add 232.4uL 29.67nM AuNP to 232.4uL 2.22uM DNA

5) React 5 min

6) Add 35.2uL sodium citrate buffer

7) React 10 minutes

8) Centrifuge 3 times for 10000rpm for 3 minutes. Redisperse all supernatants in 50mM HEPES buffer.

9) Take UV-Vis of AuNP/DNA sample and all supernatants.

Observtions

It seems as if the AuNp are falling out of solution as they are centrifuged. By the time the last cycle occurs, the solution is barely colored and there are visible purple streaks on the sides and bottom of the epitest tube. Even after vortexing the sample in HEPES buffer the nanoparticles do not go back into solution. Tomorrow I will use a nanoparticle solution that is definitely known to stay in solution after centrifugation.




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