The document describes several qualitative tests performed on hydrolyzed and unhydrolyzed DNA samples. The Dische test was expected to produce a bright blue color for hydrolyzed DNA and a brownish blue for unhydrolyzed, but expired reagents led to incorrect results. Fehling's test was predicted to give a brick red precipitate for hydrolyzed DNA hydrolysate and no reaction for unhydrolyzed. Bial's, purine, and phosphate tests were also conducted to identify breakdown products, but experimental results were not included.
The document describes several qualitative tests performed on hydrolyzed and unhydrolyzed DNA samples. The Dische test was expected to produce a bright blue color for hydrolyzed DNA and a brownish blue for unhydrolyzed, but expired reagents led to incorrect results. Fehling's test was predicted to give a brick red precipitate for hydrolyzed DNA hydrolysate and no reaction for unhydrolyzed. Bial's, purine, and phosphate tests were also conducted to identify breakdown products, but experimental results were not included.
The document describes several qualitative tests performed on hydrolyzed and unhydrolyzed DNA samples. The Dische test was expected to produce a bright blue color for hydrolyzed DNA and a brownish blue for unhydrolyzed, but expired reagents led to incorrect results. Fehling's test was predicted to give a brick red precipitate for hydrolyzed DNA hydrolysate and no reaction for unhydrolyzed. Bial's, purine, and phosphate tests were also conducted to identify breakdown products, but experimental results were not included.
The document describes several qualitative tests performed on hydrolyzed and unhydrolyzed DNA samples. The Dische test was expected to produce a bright blue color for hydrolyzed DNA and a brownish blue for unhydrolyzed, but expired reagents led to incorrect results. Fehling's test was predicted to give a brick red precipitate for hydrolyzed DNA hydrolysate and no reaction for unhydrolyzed. Bial's, purine, and phosphate tests were also conducted to identify breakdown products, but experimental results were not included.
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EXPT 4B
HYDROLYSIS OF NUCLEIC ACID
HYDROLYSIS OF DNA MOLECULE • Acid Hydrolysis (Lysis – break down) – Depurination • break down of glycosidic bonds only if pH < 3 • Products are: purine base and deoxyribose with phosphate group
– Total breakdown • break down of glycosidic bond with phosphodiester bond if pH < 2 • Extremely low pH hydrolyses DNA completely • Products are: phosphate group, the purine or pyrimidine base, and deoxyribose QUALITATIVE TESTS Dische Diphenylamine Test – for DNA • Test for the deoxyribose in DNA • Reagents – diphenylamine, Sulfuric acid • Positive result – light blue to blue depending on the amount of DNA THEORETICAL RESULTS
DISCHE TEST SAMPLES Observations
HYDROLYZED bright blue
UNHYDROLYZED brownish blue
EXPERIMENTAL RESULT RESULTS did not coincide with the expected. Error: Diphenylamine was already expired QUALITATIVE TESTS FEHLING'S Test – test for reducing sugar • Reagents: – Fehling's A: Copper sulfate solution – Fehling's B: aqueous potassium sodium tartrate and sodium hydroxide • Positive result: – formation of brick red precipitate
UNHYDROLYZED formation of brick-red ppt EXPERIMENTAL RESULTS QUALITATIVE ANALYSIS • Bial’s Test – Test for pentoses – Reagents: Orcinol, hydrochloric acid, FeCl3 – Blue or green signifies positive result THEORETICAL RESULTS
BIAL'S TEST SAMPLES Observations
Brown solution HYDROLYZED (negative for ribose)
brown solution UNHYDROLYZED (negative for ribose) EXPERIMENTAL RESULTS QUALITATIVE ANALYSIS • Test for Purine Bases – Test for the presence of Adenine and Guanine – Reagents: Silver Nitrate and Ammonium Hydroxide – Silver ion in ammoniacal solution precipitates purine bases reacting to the nitrogen of purine base THEORETICAL RESULTS
PURINE TEST SAMPLES Observations
Gelatinous white HYDROLYZED precipitate
UNHYDROLYZED brown solution
EXPERIMENTAL RESULT QUALITATIVE ANALYSIS • Test for Phosphate – Reagents: Ammonium hydroxide, nitric acid, ammonium molybdate – Yellow precipitate is the positive indicator
