User:Allison K. Alix/Notebook/Thesis Research/2013/07/25

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(Fluorescence Measurements)
(Fluorescence Measurements)
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Revision as of 14:03, 25 July 2013

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To Do

  • Repeat Silica-Coating procedure using water to dilute nanoparticles instead of PBS. PBS is likely the cause of the aggregation we have been observing.[1]
  • Repeat ThT-DNA fluorescence procedure to show that fluorescence increases with increasing number of G-quadruplexes formed.

Silica-Coating Part 1

1) Dilute 23.5nM AuNPs to 9.4nM (2mL AuNPs in 3mL water *NOTE: the water was left to assimilate to room temperature prior to dilution*)

2) Prepare 5mL 10mM MHA in water and isopropanol

3) Mix 5mL AuNPs w/ 5mL MHA

4) Stir overnight

ThT/DNA attachment

1) Prepare 100uL of the following concentrations of 4-strand and 8-strand telomeric DNA (which should generate 1 and 2 G-quadruplexes respectively)

a) 100nM

b) 300nM

c) 500nM

d) 700nM

e) 900nM

NOTE: Original Concentrations of 4 and 8-repeats are 265.6uM and 142.8uM, respectively

4-repeat

x(500uL)=(265.6uM)(10)

x = 5.31uM

[100-([x(100)]/5.31)] =

1800nM--> 33.9uL in 66.1uL PBS

1400nM--> 26.4uL in 73.6uL

1000nM--> 18.8uL in 81.2uL

600nM--> 11.3uL in 88.7uL

200nM--> 3.8uL in 96.2uL

8-repeat

142.8uM(10uL) = (x)(500uL)

x= 2.856uM

1800nM--> 63uL in 37uL PBS

1400nM--> 49uL in 51uL

1000nM--> 35uL in 65uL

600nM--> 21uL in 79uL

200nM--> 7uL in 93uL

Fluorescence Measurements

Image:OWW_pic_4_repeat.png


Image:OWW_pic_8_repeat.png







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