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

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(DNA-AuNP attachment procedure)
 
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{|{{table}} width="800"
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Absorbance Measurements of AuNP@SiO2</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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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]]
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==DNA-AuNP attachment procedure==
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*Scale down to a final solution volume of 300uL due to minimal AuNP solution.
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'''Calculating volumes of buffer, AuNP, and DNA for final solution:'''
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[sodium citrate] = 268mM
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268mM (x) = 10mM (300uL)
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x = 11.2uL sodium citrate
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(300uL-11.2uL)/2 = 144.4uL each of DNA and AuNP
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'''Procedures:'''
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1) Prepare 200uL 4.45uM DNA
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53.1uM(x) = 200uL(4.45)
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x = 16.8uL in 183.2uL 1X PBS
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2) Mix DNA with 200uL DTT in TEA
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3) React 10 minutes
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4) Extract DNA using 4 2mL aliquots of ethyl acetate
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5) Mix 144.4uL DNA with 144.4uL AuNP and 11.2uL sodium citrate
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6) React 10 min
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7) Centrifuge for 11 minutes @ 10,000rpm. Remove supernatant, redisperse in 50mM HEPES. Repeat 2 times.
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Current revision

Absorbance Measurements of AuNP@SiO2 Main project page
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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.






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