# User:Allison K. Alix/Notebook/Thesis Research/2013/09/24

(Difference between revisions)
 Revision as of 09:42, 24 September 2013 (view source) (→Preparation of Au/DNA/MB w/o Si for FCS)← Previous diff Revision as of 10:06, 24 September 2013 (view source) (→Preparation of Au/DNA/MB w/o Si for FCS)Next diff → Line 62: Line 62: Final Concentrations need to be Final Concentrations need to be - [Au] = 5.52nM + [Au] = 6.25nM [DNA] = 910nM [DNA] = 910nM + + 1) Mix 145.9uL 3.64uM thiol-DNA with 145.9uL 100mM DTT in 4% TEA. React 10 minutes + + 53.1uM(10uL) = 3.64uM(x) + + 10uL in 135.9 water + + 2) Extract DTT with 4 2mL aliquots of ethyl acetate

## Revision as of 10:06, 24 September 2013

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## Calculating [AuNP] in Au@SiO2/DNA-MB solutions

• Original [Au@SiO2] = 5nM before attaching DNA

1) Addition of 5uL APS

(1mL)(5nM)=(1.005mL)(x)

x = 4.975nM

2) Addition of 1.5mL Sulfo-SMCC

(1.005mL)(4.975nM) = (x)(2.505mL)

x = 1.996nM

3) Centrifugation, removal of supernatant, and redispersion in 530uL KP buffer

(1.996nM)(2.505mL) = x(0.530mL)

x = 9.43nM

4) Addition of 270uL DNA

(9.43nM)(530uL)=x(800uL)

x = 6.25nM

5) Addition of 13.3uL 7.61uM MB to 100uL Au@SiO2/DNA

6.25nM(100uL)=x(113.3uL)

x = 5.52nM

Dilution Factor [Au@SiO2] [DNA] [MB]
None (stock) 5.52nM 802nM 895nM
2X 2.76nM 401nM 447.5nM
10X 552pM 80.2nM 89.5nM
100X 55.2pM 8.02nM 8.95nM
1000X 5.52pM 802pM 895pM
10000X 552fM 80.2pM 89.5pM

## Preparation of Au/DNA/MB w/o Si for FCS

Final Concentrations need to be

[Au] = 6.25nM

[DNA] = 910nM

1) Mix 145.9uL 3.64uM thiol-DNA with 145.9uL 100mM DTT in 4% TEA. React 10 minutes

53.1uM(10uL) = 3.64uM(x)

10uL in 135.9 water

2) Extract DTT with 4 2mL aliquots of ethyl acetate