Difference between revisions of "User:Allison K. Alix/Notebook/Thesis Research/2013/02/19"

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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;"> ZnPPIX/DNA and ThT/DNA Sample Prep</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>}}
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|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]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
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Remake 2.0μM and 2.5μM 4-strand DNA and 2.5μM 8-strand DNA to be mixed with 3.5μM ThT (these need to be remade as they were outliers in our previous fluorescence measurements from [[User:Allison K. Alix/Notebook/Thesis Research/2013/02/12|02/12/2013]]
 
Remake 2.0μM and 2.5μM 4-strand DNA and 2.5μM 8-strand DNA to be mixed with 3.5μM ThT (these need to be remade as they were outliers in our previous fluorescence measurements from [[User:Allison K. Alix/Notebook/Thesis Research/2013/02/12|02/12/2013]]
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==Procedures==
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1) Prepare the following concentrations of DNA using calculations from [[User:Allison K. Alix/Notebook/Thesis Research/2013/02/05|02/05/2013]]. Use tris-HCL buffer to dilute.
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* 3.5μM, 3.0μM, 2.5μM, 2.0μM, 1.5μM 4-strand telomeric DNA
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* 3.5μM, 3.0μM, 2.5μM, 2.0μM, 1.5μM 8-strand telomeric DNA
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2) Mix each of the DNA samples with 250μL 5μM ZnPPIX
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 +
3) Also prepare an extra set of 2.0μM and 2.5μM 4-strand telomerics DNA and 2.5μM 8-strand DNA
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 +
4) Mix those DNA samples with 250μL 3.5μM ThT
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5) Heat all samples (total of 13) above 70°C to hybridize for 25 minutes
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6) Obtain UV-Vis and Fluorescence Spectra of all samples
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Latest revision as of 21:28, 26 September 2017

Owwnotebook icon.png ZnPPIX/DNA and ThT/DNA Sample Prep Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Objectives

Prepare samples with 5μM ZnPPIX and varying concentrations of 4-strand and 8-strand telomeric DNA

Remake 2.0μM and 2.5μM 4-strand DNA and 2.5μM 8-strand DNA to be mixed with 3.5μM ThT (these need to be remade as they were outliers in our previous fluorescence measurements from 02/12/2013

Procedures

1) Prepare the following concentrations of DNA using calculations from 02/05/2013. Use tris-HCL buffer to dilute.

  • 3.5μM, 3.0μM, 2.5μM, 2.0μM, 1.5μM 4-strand telomeric DNA
  • 3.5μM, 3.0μM, 2.5μM, 2.0μM, 1.5μM 8-strand telomeric DNA

2) Mix each of the DNA samples with 250μL 5μM ZnPPIX

3) Also prepare an extra set of 2.0μM and 2.5μM 4-strand telomerics DNA and 2.5μM 8-strand DNA

4) Mix those DNA samples with 250μL 3.5μM ThT

5) Heat all samples (total of 13) above 70°C to hybridize for 25 minutes

6) Obtain UV-Vis and Fluorescence Spectra of all samples