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

From OpenWetWare

< User:Allison K. Alix | Notebook | Thesis Research | 2013 | 07(Difference between revisions)
Jump to: navigation, search
(DNA-AuNP attachment procedure)
Current revision (09:19, 17 July 2013) (view source)
(DNA-AuNP attachment procedure)
 
Line 48: Line 48:
6) React 10 min
6) React 10 min
-
7) Centrifuge for 10 minutes @ 11,000rpm. Remove supernatant, redisperse in 50mM HEPES. Repeat 2 times.
+
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.






Personal tools