This preview has intentionally blurred sections. Sign up to view the full version.View Full Document
Unformatted text preview: My-Linh TranSect: 0104Exp: 04/26/11Quantitative Determination of DNA-Ligand Binding ConstantGoal: Use fluorescence intensity measurements to find the binding constant with calf-thymus DNA for kinetics by studying DAPI complex. Studying kinetic measurements can provide information on the properties of the molecule: number of nucleotides per bound ligand. Compounds: 0.02 M NaOH solution in distilled water 0.02 M solution of sodium dihydrogen phosphate in distilled water 4’, 6-diamidino-2-phenylindole (DAPI), 1.0x10-4M stock solution in ethanol (keep on ice)Calf-thymus DNA, 50 µg/ml solution in 0.01 M phosphate buffer (pH=7.4)WavelengthsExcitation: 360 nm Emission: 450 nm General Plan for the Lab:Prepare stock solution; 0.02 M of NaOH and 0.02 M sodium dihydrogen phosphate will be available under the fume hood. Prepare 0.01 M of phosphate buffer by mixing 40 ml of 0.02 M NaOH and 50 ml of 0.02 sodium dihydrogen phosphate and diluting with 100 ml of DI water in a 250 container (beaker, flask). ml of DI water in a 250 container (beaker, flask)....
View Full Document
- Spring '08
- DNA, Sodium hydroxide, nucleotide, Phosphate, DAPI