User:Allison K. Alix/Notebook/Thesis Research/2013/09/24: Difference between revisions
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x = 6.25nM | 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 | |||
{| class="wikitable" | |||
|- | |||
! Dilution Factor !! [Au@SiO<sub>2</sub>] !! [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 | |||
3) Prepare 100uL 12.5nM AuNP | |||
23.5(x) = 12.5(100uL) | |||
x = 53.2uL in 46.8uL water | |||
4) mix 20uL DNA with 20uL AuNP and 5uL 268mM sodium citrate. React 10 minutes. | |||
5) Centrifuge 7 minutes at 10000rpm. | |||
6) Add 6uL MB heat for 25 min at 70 C | |||
==Notes== | |||
FCS Measurements for the Au@Si/DNA-MB samples were run twice each for 7 minutes at a time. Looking at the N values, we have determined that the range for detection is between the 50X and 100X diluted samples (AuNP concentrations of 110pM and 55.2pM respectively.) The Au/DNA-MB samples were not run for this day as they required a few hours to cool to room temperature. This being said, they will also be run in this same 50X-100X diltuion range. | |||
Revision as of 07:34, 25 September 2013
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Calculating [AuNP] in Au@SiO2/DNA-MB solutions
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
Preparation of Au/DNA/MB w/o Si for FCSFinal 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 3) Prepare 100uL 12.5nM AuNP 23.5(x) = 12.5(100uL) x = 53.2uL in 46.8uL water 4) mix 20uL DNA with 20uL AuNP and 5uL 268mM sodium citrate. React 10 minutes. 5) Centrifuge 7 minutes at 10000rpm. 6) Add 6uL MB heat for 25 min at 70 C NotesFCS Measurements for the Au@Si/DNA-MB samples were run twice each for 7 minutes at a time. Looking at the N values, we have determined that the range for detection is between the 50X and 100X diluted samples (AuNP concentrations of 110pM and 55.2pM respectively.) The Au/DNA-MB samples were not run for this day as they required a few hours to cool to room temperature. This being said, they will also be run in this same 50X-100X diltuion range.
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