User:Allison K. Alix/Notebook/Thesis Research/2013/12/16: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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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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Latest revision as of 23:37, 26 September 2017
Project name | Main project page Previous entry Next entry |
NEW Telomeric DNAthiol-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 Attachmentoriginal [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 |