User:Allison K. Alix/Notebook/Thesis Research/2013/01/29

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(Procedures)
(Procedures)
Line 23: Line 23:
x = '''2.656 μL''' in 97.344 μL H<sub>2</sub>O
x = '''2.656 μL''' in 97.344 μL H<sub>2</sub>O
 +
2. Prepare 1 μM 8 telomeric repeat DNA
2. Prepare 1 μM 8 telomeric repeat DNA
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- Add 1 N HCl (~8 drops) to obtain pH ~ 7.25
- Add 1 N HCl (~8 drops) to obtain pH ~ 7.25
 +
2. prepare 3.5 μM ThT
2. prepare 3.5 μM ThT
Line 46: Line 48:
x = '''0.01mL''' in 9.99mL H<sub>2</sub>O
x = '''0.01mL''' in 9.99mL H<sub>2</sub>O
-
3. Prepare 3.5μM, 3μM, 2.5μM, 2.0μM, and 1.5μM solutions of 4 telomeric repeat DNA
 
 +
3. Prepare 3.5μM, 3μM, 2.5μM, 2.0μM, and 1.5μM solutions of 4 telomeric repeat DNA
a) (265.6μM)(x) = (3.5μM) (250μL)
a) (265.6μM)(x) = (3.5μM) (250μL)
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::x= '''2.82μL''' in 247.18μL tris-HCl buffer
::x= '''2.82μL''' in 247.18μL tris-HCl buffer
 +
c) (265.6μM)(x) = (2.5μM) (250μL)
c) (265.6μM)(x) = (2.5μM) (250μL)

Revision as of 20:22, 4 February 2013

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Objectives

-Repeat ZnPPIX and DNA hybridizations in 5:1 ratios instead of 1:1

-Hybridize ThT and DNA strands

Procedures

ZnPPIX:

5μM ZnPPIX already prepared

1. Prepare 1 μM 4 telomeric repeat DNA:

265.6μM (x) = 1μM (100μL)

x = 2.656 μL in 97.344 μL H2O


2. Prepare 1 μM 8 telomeric repeat DNA

142.8μM (x) = 1μM (100μL)

x = 1.428 μL in 98.572 μL H2O

ThT:

1. prepare 5mM tris-HCl buffer solution

121.4 g/mol x 5mmol/L x 1mol/1000mmol x 50 mL x 1L/1000mL = 0.030285 g tris in 50 mL H2O

- Add 1 N HCl (~8 drops) to obtain pH ~ 7.25


2. prepare 3.5 μM ThT

318.86 g/mol x 3.5mmol/L x 1mol/1000mmol x 10mL x 1L/1000mL = 0.0111601 g in 10mL H2O -> 3.5mM

(3.5 x 10^-3 M) (x) = (3.5 x 10^-6 M) (10mL)

x = 0.01mL in 9.99mL H2O


3. Prepare 3.5μM, 3μM, 2.5μM, 2.0μM, and 1.5μM solutions of 4 telomeric repeat DNA

a) (265.6μM)(x) = (3.5μM) (250μL)

x= 3.29μL in 246.71μL tris-HCl buffer


b) (265.6μM)(x) = (3.0μM) (250μL)

x= 2.82μL in 247.18μL tris-HCl buffer


c) (265.6μM)(x) = (2.5μM) (250μL)

x= 2.35μL in 247.65μL tris-HCl buffer


d) (265.6μM)(x) = (2.0μM) (250μL)

x= 1.88μL in 248.12μL tris-HCl buffer


e) (265.6μM)(x) = (1.5μM) (250μL)

x= 1.41μL in 248.59μL tris-HCl buffer




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