# User:Allison K. Alix/Notebook/Thesis Research/2013/07/17

(Difference between revisions)
 Revision as of 15:07, 16 July 2013 (view source)← Previous diff Current revision (10:19, 17 July 2013) (view source) (→DNA-AuNP attachment procedure) (2 intermediate revisions not shown.) Line 15: Line 15: * Obtain UV-Vis Data for DNA/AuNP samples and perform calculations following steps from [[User:Allison K. Alix/Notebook/Thesis Research/2013/07/16]] * Obtain UV-Vis Data for DNA/AuNP samples and perform calculations following steps from [[User:Allison K. Alix/Notebook/Thesis Research/2013/07/16]] + + ==DNA-AuNP attachment procedure== + + *Scale down to a final solution volume of 300uL due to minimal AuNP solution. + + '''Calculating volumes of buffer, AuNP, and DNA for final solution:''' + + [sodium citrate] = 268mM + + 268mM (x) = 10mM (300uL) + + x = 11.2uL sodium citrate + + (300uL-11.2uL)/2 = 144.4uL each of DNA and AuNP + + '''Procedures:''' + + 1) Prepare 200uL 4.45uM DNA + + 53.1uM(x) = 200uL(4.45) + + x = 16.8uL in 183.2uL 1X PBS + + 2) Mix DNA with 200uL DTT in TEA + + 3) React 10 minutes + + 4) Extract DNA using 4 2mL aliquots of ethyl acetate + + 5) Mix 144.4uL DNA with 144.4uL AuNP and 11.2uL sodium citrate + + 6) React 10 min + + 7) Centrifuge for 11 minutes @ 10,000rpm. Remove supernatant, redisperse in 50mM HEPES. Repeat 2 times. + + + + + + +

## Current revision

Absorbance Measurements of AuNP@SiO2 Main project page
Previous entry      Next entry

## To Do

• Take absorbance spectra of AuNP@SiO2 samples
• Determine which concentrations are suitable for TEM imaging (Thursday)
• Repeat DNA/AuNP extraction procedure using optimum centrifugation speed and time (10,000 rpm for 11 minutes)

## DNA-AuNP attachment procedure

• Scale down to a final solution volume of 300uL due to minimal AuNP solution.

Calculating volumes of buffer, AuNP, and DNA for final solution:

[sodium citrate] = 268mM

268mM (x) = 10mM (300uL)

x = 11.2uL sodium citrate

(300uL-11.2uL)/2 = 144.4uL each of DNA and AuNP

Procedures:

1) Prepare 200uL 4.45uM DNA

53.1uM(x) = 200uL(4.45)

x = 16.8uL in 183.2uL 1X PBS

2) Mix DNA with 200uL DTT in TEA

3) React 10 minutes

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

5) Mix 144.4uL DNA with 144.4uL AuNP and 11.2uL sodium citrate

6) React 10 min

7) Centrifuge for 11 minutes @ 10,000rpm. Remove supernatant, redisperse in 50mM HEPES. Repeat 2 times.