User:Allison K. Alix/Notebook/Thesis Research/2013/12/16

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(DNA Attachment)
Current revision (13:26, 16 December 2013) (view source)
(DNA Attachment)
 
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attach DNA to AuNP in 75:1 ratio
attach DNA to AuNP in 75:1 ratio
 +
Final concentrations should be 881.25nM DNA and 11.75nM AuNP
 +
1) Prepare 250uL 3.525uM thiol-DNA
 +
63.1uM (x) = 250uL(3.525uM)
 +
 +
x = 13.97uL 63.1 uM thiol-DNA in 236.03uL 1X PBS
 +
 +
2) React 10 minutes and extract DNA using ethyl acetate
 +
 +
3) Mix DNA with sodium citrate to change pH (THIS MUST BE DONE PRIOR TO ADDING AUNP!!!)
 +
 +
268mM sodium citate buffer
 +
 +
268mM (x) = 10mM(500uL)
 +
 +
x = 18.7uL
 +
 +
500uL-18.7uL = 481.3uL
 +
 +
481.3uL/2 = 240.65uL each of AuNP and DNA
 +
 +
4) Add 240.65uL AuNP
 +
 +
5) React 10 minutes
 +
 +
6) Centrifuge for 4 minutes at 7000rpm
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NEW Telomeric DNA

thiol-8 repeat GTTAGGG

Tm=69.2°C

MW=19,007.5

38.2OD= 62.9nm = 1.20mg

1.20mg x mol/19,007.5g x 1g/1000mg = 6.31x10-8mol

dilute in 1mL 1X PBS

6.31x10-8mol / 1mL x 1000mL/1L = 6.31x10-5M

6.31x10-5M x 106μM/1M = 63.1μM

DNA Attachment

original [AuNP] = 23.5nM

original [DNA] 63.1uM

attach DNA to AuNP in 75:1 ratio

Final concentrations should be 881.25nM DNA and 11.75nM AuNP

1) Prepare 250uL 3.525uM thiol-DNA

63.1uM (x) = 250uL(3.525uM)

x = 13.97uL 63.1 uM thiol-DNA in 236.03uL 1X PBS

2) React 10 minutes and extract DNA using ethyl acetate

3) Mix DNA with sodium citrate to change pH (THIS MUST BE DONE PRIOR TO ADDING AUNP!!!)

268mM sodium citate buffer

268mM (x) = 10mM(500uL)

x = 18.7uL

500uL-18.7uL = 481.3uL

481.3uL/2 = 240.65uL each of AuNP and DNA

4) Add 240.65uL AuNP

5) React 10 minutes

6) Centrifuge for 4 minutes at 7000rpm


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