DNA is stable to bases, these is one of thefeatures that distinguish DNA from RNA. Strong acids athigh temperatures are capable of breaking down DNAmolecules onto its basic components, at theseconditions both of the phosphate ester bond and the N-glycosides bends between the Deoxyribose, the Purinesand the pyrimidine bases.
Compounds Tested; Samples used and DevicesusedDNA isolation from Onion
The following were used for the isolation of DNA; 2 White/yellow Onions, Homogenizing Solutions;5% SDS( Sodium Dodecyl Sulfate), 0.15 M NaCl, 0.15 MNa
and 0.001 M EDTA, Ice-Cold 95% ethanol.
Ultraviolet Measurement of DNA
The Following were used in the Measurements:Quartz Cuvettes, UV-VIS Spectrophotometer.
The Following were used in the Acid Hydrolysis:DNA Sample, 1M HCl
Test for Deoxyribose
The following were used in the test: 3.5mlDiphenylamine reagent, 1.5ml Hydrolyzed DNAsolution, 0.5 ml Standard DNA, Hot water Bath
Test for Phosphate
The Following were used in the Test forphosphates: 1ml conc. H
, 1ml Nucleic Acid Solution,1ml Standard Phosphate solution, 0.5ml conc.HNO
O, 1ml Molybdate Solution, 10ml water.
Test for Purines
The Following were used in the Murexide Test:5-10GTT Nucleic Acid solution, HNO
,10% KOH, water
Test for Pyrimidines
Lastly the following were used in the test: 0.5mlNucleic Acid solution, Bromine Water, BariumHydroxide
Isolation of DNA from Onion.
A 50 ml Homogenizing solution is then preparedin a 125 ml Erlenmeyer flask and was heated in a waterbath until the solution reached 60 deg C, 25 g mincedonion was added to the preheated water bath. A 1.5gpapain was then added and was then kept for 10minutes. After heating place the Erlenmeyer to an icebath for about 5 minutes, after the cooling process,place the solution to in a beaker to be homogenized;after the homogenization process the homogenate wasthen filtered using a cheese cloth into a 250ml beakerand cool it after wards. The cooled solution was thenadded with 15-20ml of ice-cold ethanol, place it on thesides and allow it to drip downwards, a clear layer of ethanol should form on top of the solution, DNA is notsoluble in ice-cold ethanol, when ethanol is added tothe mixture, all the components in the solution exceptfor the DNA stay in the solution, while the DNAprecipitates out. Let the mixture stand for 3-5 minuteswithout disturbing, the DNA can be perceived by awhite precipitate/ strings. Spool the Formed DNA byusing a pre-bent glass pipette, the obtained specimenwas then added with TE buffer solution.
Ultraviolet Measurement of Isolated DNA
The obtained DNA solution (0.5ml) is thenadded to a 4.5 SSC or TE buffer solution. It is thetransferred to a quartz cuvette to determine theabsorbance at the range of 260nm-280nm. Theobtained results are then calculated for the ratio todetermine the concentrations of protein and nucleicacid.
Mix the DNA sample with 1M HCl, then heat itup to 100deg C for 60 minutes with occasional agitation,cover the tubes with marbles and placed under arunning water to neutralize the temperature and add