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

(Difference between revisions)
 Revision as of 20:12, 4 February 2013 (view source) (→Procedures)← Previous diff Revision as of 20:18, 4 February 2013 (view source) (→Procedures)Next diff → Line 34: Line 34: 1. prepare 5mM tris-HCl buffer solution 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 + 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 - Add 1 N HCl (~8 drops) to obtain pH ~ 7.25 Line 40: Line 40: 2. prepare 3.5 μM ThT 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 + 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) (3.5 x 10^-3 M) (x) = (3.5 x 10^-6 M) (10mL) - x = 0.01mL in 9.99mL H2O + 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

## Revision as of 20:18, 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:

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