User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/02/26

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February 26, 2013

Hybridization of molecular beacon and complementary strand at varying concentrations

MB:CS (MB at 500pM)

(7.3x10^-6M)V=(3x10^-6)(10μL) 4.1μL MB in 5.9μL buffer <br.> (2.8x10^-5)V=(3x10^-6M)(10μL) 1.1μL CS in 8.9μL buffer <br.> (3x10^-6)V=(500x10^-9)(50μL) 8.33μL in 41.67μL <br.>

  • 1:1 (1μL MB:1μL CS) - 0.5μM MB:0.5μM CS
  • 1:5 (1μL MB:5μL CS) - 0.2μM MB:0.8μM CS
  • 1:10 (1μL MB:10μL CS) - 0.09μM MB:0.91μM CS

<br.>

MB:CS (CS at 500pM)

(7.3x10^-6)V=(5x10^-6)(5μL) 3.42μL MB in 1.57μL buffer <br.> 0.36μL CS in 9.64μL buffer

  • 1:1 (1μL MB:1μL CS) - 0.5μM MB:0.5μM CS
  • 5:1 (5μL MB:1μL CS) - 0.8μM MB:0.2μM CS
  • 10:1 (10μL MB:1μL CS) - 0.91μM MB:0.09μM CS
  • 50:1 (50μL MB:1μL CS) - 0.98μM MB:0.02μM CS

<br.>

Make 1nm, 500pM, and 100pM samples for each ratio

1:1

  1. 50nM: (500nM)V=(50nM)(50μL)
    5μL 500nM solution in 45μL buffer
  1. 1nM: (50nM)V=(1nM)(50μL)
     1μL 50nM solution in 49μL buffer
  1. 500pM (0.5nM): (1nM)V=(0.5nM)(50μL)
     25μL 1nM solution in 25μL buffer
  1. 100pM (0.1nM): (0.5nM)V=(0.1nM)(50μL)
     10μL 0.5nM solution in 40μL buffer

1:5, 1:10, 5:1, 10:1, and 50:1 calculations done in the same way



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