SHEA HIGH SCHOOL July 13-17, 2009 Ms. La Roche Cesar Luzon, Janice Miranda, Maldine Santos, Estefania Tabares.

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Presentation transcript:

SHEA HIGH SCHOOL July 13-17, 2009 Ms. La Roche Cesar Luzon, Janice Miranda, Maldine Santos, Estefania Tabares.

Formula : CH 3 P(C 6 H 5 ) 4 Mol WT : Synonyms : Bromure de tétraphénylphosphonium; Bromuro de tetrafenilfosfonio; Classification : ORGANOPHOSPHINES. ORGANOPHOSPHINES Physical state : white to off-white crystalline powder. Melting Point : C Solubility in water : Soluble. Stability: Stable under ordinary conditions.

Well 1: 10.0 uL Linearized Plasmid DNA. Well 2: 10.0 uL uncut Plasmid DNA Well 3: 10.0uL linearized plasmid DNA w B-A. Well 4: 10.0 uL uncut plasmid DNA w B-A. Well 5: 5.0 uL Kb ladder

Electrophoresis Distance (mm)Base parirs 1923, , , , , , DistanceBase pairs DistanceBase pairs E-linearEB-linear plasmid& TPP Observations : both the linear and the uncut plasmid with TPP travelled 1mm further showing the small molecule bonded with the DNA molecule. X-uncutXB-uncut plasmid& TPP

Temp.CºAbsorbance∆T∆ATave∆A/∆T DNA Temp. CºAbsorbance∆TA∆Tave∆A/∆T DNA+orbizine

Salmon DNA +TPP

MAJOR GROOVEMINOR GROOVEINTERCALATING

TESTORIENTATIONMAJOR GROVE MINOR GROVE INTERCALATIN G DNA and TPP X DNA and TPP Y DNA and TPP Z

Temp. C°Absorbance (nm)Column1Column2Column3Column4 DNAΔTΔTΔAΔATaveΔA/ΔT Temp.C°Absorbance (nm)Column1Column2Column3Column4 DNA+TPP ΔTΔAΔATaveΔA/ΔT Absorbance at 260 nm

Hyperchem analyses led to the prediction that tetraphenylphosphonium Bromide would bond to DNA. The experimental data shows a shift in the melting curve when TPP is added, agreeing with this prediction No significant changes were made in the experimental procedures. Light banding in electrophoresis gels suggest using a larger amount of the Samples. Practice pipetting into wells is suggested. Repeated trials of both experimental procedures are recommended for further research. The melting point of Salmon DNA was 70 ⁰ C on the Smart Spec and 59 ⁰ C on the Cary spectrophotometer. The difference in the melting point could be due to the fluctuating temperatures with the Smart Spec Plus. The melting point of the Salmon DNA with the small molecule TPP: Cary - appears to be 49 ⁰ C. There are three other indications of melting in different phases. Smart Spec – points to two melting points, 68 ⁰ C and 83 ⁰ C.