User:Allison K. Alix/Notebook/Thesis Research/2013/01/29
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Objectives-Repeat ZnPPIX and DNA hybridizations in 5:1 ratios instead of 1:1 -Hybridize ThT and DNA strands ProceduresZnPPIX:
265.6μM (x) = 1μM (100μL) x = 2.656 μL in 97.344 μL H2O
142.8μM (x) = 1μM (100μL) x = 1.428 μL in 98.572 μL H2O 3) Mix 100μL ZnPPIx with 100μL of each DNA sequences
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
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
a) (265.6μM)(x) = (3.5μM) (250μL)
b) (265.6μM)(x) = (3.0μM) (250μL)
c) (265.6μM)(x) = (2.5μM) (250μL)
d) (265.6μM)(x) = (2.0μM) (250μL)
e) (265.6μM)(x) = (1.5μM) (250μL)
a) (142.8μM) (x) = (3.5μM)(250μL)
b) (142.8μM) (x) = (3.0μM)(250μL)
c) (142.8μM) (x) = (2.5μM)(250μL)
d) (142.8μM) (x) = (2.0μM)(250μL)
e) (142.8μM) (x) = (1.5μM)(250μL)
5) Mix 100μL ThT with each of the prepared DNA samples in all concentrations
1) Heat all hybridization samples for ~20min at ~76°C 2) Take absorbance and fluorescence of all (to be done next week)
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