User:Allison K. Alix/Notebook/Thesis Research/2013/11/19

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(Sample Preparation)
Current revision (11:55, 20 November 2013) (view source)
(Sample Preparation)
 
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4) Cool to room temperature
4) Cool to room temperature
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==Notes==
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The purpose of this analysis is to observe if fluorescence changes with changing concentrations of Au/DNA. Where as before one Au/DNA/MB solution was prepared and subsequent dilutions were made from that one solution, this procedure first prepares the dilutions and attaches the MB to each separate solution. The higher concentrations of Au/DNA have more binding sites for the beacon, so they are expected to bind fully. The lower concentrations should also bind fully with some closed MB in solution.

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To Do

  • Vary concentrations of AuNP and DNA.
  • Add the same amount of MB to all samples

Sample Preparation

1) Prepare the following dilutions of AuNP/DNA:

original concentration of AuNP: 290pM

20X 14.5pM

40X 7.25pM

60X 4.83pM

80X 3.625pM

100X 2.9pM

120X 2.4167pM

140X 2.07pM

160X 1.8125pM

180X 1.61pM

200X 1.45pM

240X 1.208pM


2) Add 1uL 7.61uM Molecular Beacon to each solution.

3) Heat at ~78°C for 30 minutes

4) Cool to room temperature

Notes

The purpose of this analysis is to observe if fluorescence changes with changing concentrations of Au/DNA. Where as before one Au/DNA/MB solution was prepared and subsequent dilutions were made from that one solution, this procedure first prepares the dilutions and attaches the MB to each separate solution. The higher concentrations of Au/DNA have more binding sites for the beacon, so they are expected to bind fully. The lower concentrations should also bind fully with some closed MB in solution.




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