User:Allison K. Alix/Notebook/Thesis Research/2013/07/17: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Absorbance Measurements of AuNP@SiO2</span> | ||
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* 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. | |||
Revision as of 07:19, 17 July 2013
 Absorbance Measurements of AuNP@SiO2
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To Do
DNA-AuNP attachment procedure
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.
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