The purpose of this experiment is to examine the extraction of the DNA material that was extracted from the Chromosomal DNA from E coli in laboratory 9. DNase is used to help extract the Sugar phosphate backbone of the DNA. DNase should only cleave DNA not RNA but DNase can classify contaminated RNA from chromosomal DNA.
Methods A. preparation :E. coli was used to extract the DNA as instructed in laboratory 9 to use in the experiment on analyzing in this experiment. There was 500 μL of the DNA material extracted from laboratory 9 to use for testing.
B. enzymatic digestion: In this experiment there was a total of 8 tubes prepared with the final volume of 20 μL. Starting with test tube 1, 2, and 4, there was 10μL of water, 10 μL of nucleic acid, and 2 μL of buffer solution (10X) are all added. In test tube 3,5, and 6 there was 5 μL of water, 10μL of nucleic acid, 2μL of buffer and 3 μL of DNase added. Although in test tube 6 the DNase is heated making in inactive. All of the other DNase are active and can consume the DNA present in the test tubes. In test 7 the same content …show more content…
In this experiment, agarose gel electrophoresis was used in to analysis the DNA that was extracted. The nucleic acid is at PH of 8 which allows the material to move down the electric field. The nucleic acid moves from positive to negative electric field. The ETBR is used to make the nucleic acid visible in the fluorescent light and the binding to the DNA. The “stop buffer” which is EDTA and bromophenol blue. The Bromophenol helped make the progress of making the DNA visible when it is injected in the gel and the EDTA increases the density of the material. The EDTA also helps stoop the DNase reaction If the DNA was fully digested by DNase, it was indicated by the lack of a band. The DNase is was starts the reaction to occur it is the last component added to each test tube that is required
Methods A. preparation :E. coli was used to extract the DNA as instructed in laboratory 9 to use in the experiment on analyzing in this experiment. There was 500 μL of the DNA material extracted from laboratory 9 to use for testing.
B. enzymatic digestion: In this experiment there was a total of 8 tubes prepared with the final volume of 20 μL. Starting with test tube 1, 2, and 4, there was 10μL of water, 10 μL of nucleic acid, and 2 μL of buffer solution (10X) are all added. In test tube 3,5, and 6 there was 5 μL of water, 10μL of nucleic acid, 2μL of buffer and 3 μL of DNase added. Although in test tube 6 the DNase is heated making in inactive. All of the other DNase are active and can consume the DNA present in the test tubes. In test 7 the same content …show more content…
In this experiment, agarose gel electrophoresis was used in to analysis the DNA that was extracted. The nucleic acid is at PH of 8 which allows the material to move down the electric field. The nucleic acid moves from positive to negative electric field. The ETBR is used to make the nucleic acid visible in the fluorescent light and the binding to the DNA. The “stop buffer” which is EDTA and bromophenol blue. The Bromophenol helped make the progress of making the DNA visible when it is injected in the gel and the EDTA increases the density of the material. The EDTA also helps stoop the DNase reaction If the DNA was fully digested by DNase, it was indicated by the lack of a band. The DNase is was starts the reaction to occur it is the last component added to each test tube that is required