User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/09/04: Difference between revisions

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"Self Assembly of the ZnPPIX/G-Quadruplex Complex" in [http://pubs.acs.org/doi/abs/10.1021/ac300348v Fluorescence Detection of DNA, Adenosine-5′-Triphosphate (ATP), and Telomerase Activity by Zinc(II)-Protoporphyrin IX/G-Quadruplex Labels]<br.>
"Self Assembly of the ZnPPIX/G-Quadruplex Complex" in [http://pubs.acs.org/doi/abs/10.1021/ac300348v Fluorescence Detection of DNA, Adenosine-5′-Triphosphate (ATP), and Telomerase Activity by Zinc(II)-Protoporphyrin IX/G-Quadruplex Labels]<br.>
#100uL 3uM 4-strand DNA- in place of telomerase solution
#100uL 3uM 4-strand DNA- in place of telomerase solution
#*100uL 50mM KCl and 5uM ZnPPIX (to ensure excess for G-quadruplex formation)
#*100uL 50mM KCl and 5uM ZnPPIX (to ensure excess for 1 G-quadruplex formation)
#*Allow to react for 10 minutes
#*Allow to react for 10 minutes
#100uL 3uM 8-strand DNA- in place of telomerase solution
#100uL 3uM 8-strand DNA- in place of telomerase solution
#*100uL 50mM KCl and 10uM ZnPPIX (to ensure excess for G-quadruplex formation)
#*100uL 50mM KCl and 10uM ZnPPIX (to ensure excess for 2 G-quadruplex formations)
#*Allow to react from 10 minutes
#*Allow to react from 10 minutes


===Modifying Fluorescence Samples from [http://openwetware.org/wiki/User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/06/05 06/05/2013]===
===Modifying Fluorescence Samples from [http://openwetware.org/wiki/User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/06/05 06/05/2013]===
#2.22uM ZnPPIX/1.11uM 4-strand DNA (to ensure excess for G-quadruplex formation)
#3.33uM ZnPPIX/1.11uM 8-strand DNA (to ensure excess for G-quadruplex formation)


#2.22uM ZnPPIX/1.11uM 4-strand DNA (to ensure excess for 1 G-quadruplex formation)
#3.33uM ZnPPIX/1.11uM 8-strand DNA (to ensure excess for 2 G-quadruplex formations)
===Fluorescence Spectra===
[[Image:2013_0904_ZnPPIX_fluor.PNG]]
===Notes===
*No signal observed for any of the samples.  The raw data was found to be exactly the same for each sample.  UV-vis and fluorescence measurements will be taken again next time.


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Revision as of 07:28, 4 December 2013

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September 4, 2013

Zinc protophorphyrin (IX) sample preparation

Replicating Fluorescence Samples from 06/05/2013

  1. 2.22uM ZnPPIX
  2. 2.22uM ZnPPIX/1.11uM DNA-SH
  3. 1.934uM ZnPPIX/0.94uM DNA-SH/11.75uM AuNPs
  4. 2.22uM ZnPPIX/11.75uM AuNPs

Replicating Fluorescence Procedure from Publication

"Self Assembly of the ZnPPIX/G-Quadruplex Complex" in Fluorescence Detection of DNA, Adenosine-5′-Triphosphate (ATP), and Telomerase Activity by Zinc(II)-Protoporphyrin IX/G-Quadruplex Labels<br.>

  1. 100uL 3uM 4-strand DNA- in place of telomerase solution
    • 100uL 50mM KCl and 5uM ZnPPIX (to ensure excess for 1 G-quadruplex formation)
    • Allow to react for 10 minutes
  2. 100uL 3uM 8-strand DNA- in place of telomerase solution
    • 100uL 50mM KCl and 10uM ZnPPIX (to ensure excess for 2 G-quadruplex formations)
    • Allow to react from 10 minutes

Modifying Fluorescence Samples from 06/05/2013

  1. 2.22uM ZnPPIX/1.11uM 4-strand DNA (to ensure excess for 1 G-quadruplex formation)
  2. 3.33uM ZnPPIX/1.11uM 8-strand DNA (to ensure excess for 2 G-quadruplex formations)

Fluorescence Spectra

Notes

  • No signal observed for any of the samples. The raw data was found to be exactly the same for each sample. UV-vis and fluorescence measurements will be taken again next time.


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