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

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===Hybridization of molecular beacon and complementary strand at varying concentrations===
===Hybridization of molecular beacon and complementary strand at varying concentrations===
====MB:CS (MB at 500pM)====
====MB:CS (MB at 500pM)====
-
(7.3x10^-6M)V=(3x10^-6)(10μL)
+
(7.3x10^-6M)V=(3x10^-6)(10μL)<br.>
4.1μL MB in 5.9μL buffer
4.1μL MB in 5.9μL buffer
<br.>
<br.>
-
(2.8x10^-5)V=(3x10^-6M)(10μL)
+
(2.8x10^-5)V=(3x10^-6M)(10μL)<br.>
1.1μL CS in 8.9μL buffer
1.1μL CS in 8.9μL buffer
<br.>
<br.>
-
(3x10^-6)V=(500x10^-9)(50μL)
+
(3x10^-6)V=(500x10^-9)(50μL)<br.>
8.33μL in 41.67μL  
8.33μL in 41.67μL  
<br.>
<br.>
Line 32: Line 32:
*50:1 (50μL MB:1μL CS) - 0.98μM MB:0.02μM CS
*50:1 (50μL MB:1μL CS) - 0.98μM MB:0.02μM CS
<br.>
<br.>
-
Make 1nm, 500pM, and 100pM samples for each ratio
+
=====Make 1nm, 500pM, and 100pM samples for each ratio=====
 +
1:1
 +
*50nM: (500nM)V=(50nM)(50μL)
 +
5μL 500nM solution in 45μL buffer
 +
*1nM: (50nM)V=(1nM)(50μL)
 +
1μL 50nM solution in 49μL buffer
 +
*500pM (0.5nM): (1nM)V=(0.5nM)(50μL)
 +
25μL 1nM solution in 25μL buffer
 +
*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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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

  • 50nM: (500nM)V=(50nM)(50μL)

5μL 500nM solution in 45μL buffer

  • 1nM: (50nM)V=(1nM)(50μL)

1μL 50nM solution in 49μL buffer

  • 500pM (0.5nM): (1nM)V=(0.5nM)(50μL)

25μL 1nM solution in 25μL buffer

  • 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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