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

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
(2.8x10^-5)V=(3x10^-6M)(10μL) 1.1μL CS in 8.9μL buffer
(3x10^-6)V=(500x10^-9)(50μL) 8.33μL in 41.67μL

• 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

MB:CS (CS at 500pM)

(7.3x10^-6)V=(5x10^-6)(5μL) 3.42μL MB in 1.57μL buffer
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

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