User:Allison K. Alix/Notebook/Thesis Research/2013/02/19: 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;"> 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> </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> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | ||
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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]] | ||
==Procedures== | |||
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. | |||
* 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 | |||
Revision as of 10:38, 19 February 2013
ZnPPIX/DNA and ThT/DNA Sample Prep | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
ObjectivesPrepare 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 Procedures1) Prepare the following concentrations of DNA using calculations from 02/05/2013. Use tris-HCL buffer to dilute.
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
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