DNA Recombination
Recombinant DNA Report Our final annotated gel image sums up the successful experiments we performed over the course of 8 weeks. The image will be referred to throughout the report:
Lane 1: 10 µL of ladder.
Lane 2: 20 µL of a pAMP- EcoRI/HindIII double digestion. Within the double digestion, one can find 8 µL of pAMP, 1 µL of the EcoRI enzyme, 1 µL of the HindIII enzyme, 5 µL of 10x Buffer 2.1, and 35 µL of water. A total volume of 50 µL was present in our 2 mL tube. Again, only 20 µL were used for the gel electrophoresis.
Lane 3: 7 µL of a pKAN- EcoRI/HindIII double digestion. Within the double digestion, one can find 38 µL of pKAN, 3 µL of the EcoRI enzyme, 3 µL of the HindIII enzyme, 6 µL of 10x Buffer 2.1, and 10 µL of water. A total volume of 60 µL was present in our 2 mL tube. Again, only 7 µL were used for the gel electrophoresis.
Lane 4: 12 µL sample of a pAMP purified sample. Within the sample, one can find 5 µL of pAMP, 5 µL of water, and 2 µL of loading dye.
Lane 5: 12 µL sample of a pKAN purified sample. Within the sample, one can find 5 µL of pKAN, 5 µL of water, and 2 µL of loading dye.
Lane 6: 5 µL sample of a pKAMP (pAMP/pKAN)- EcoRI/HindIII double digestion. Within the double digestion, one can find 2 µL of pKAMP, 1 µL of the EcoRI enzyme, 1 µL of the HindIII enzyme, 3 µL of 10x Buffer 2.1, and 23 µL of water. A total volume of 30 µL was present in our 2 mL tube. Again, only 5 µL were used for the gel electrophoresis.
Lane 7: 10 µL of ladder. We began our goal of combining DNA by obtaining the desired samples of plasmid DNA from a liquid bacteria culture. Plasmids are circular pieces of DNA, which can be found in bacteria. Plasmids are often used in the manipulation of genes. After carefully following a mini-prep procedure, we were left with samples of pAMP and pKAN plasmids. pAMP is a bacterial plasmid, which contains the gene ampR. The ampR gene codes for a protein that breaks down the
Lane 1: 10 µL of ladder.
Lane 2: 20 µL of a pAMP- EcoRI/HindIII double digestion. Within the double digestion, one can find 8 µL of pAMP, 1 µL of the EcoRI enzyme, 1 µL of the HindIII enzyme, 5 µL of 10x Buffer 2.1, and 35 µL of water. A total volume of 50 µL was present in our 2 mL tube. Again, only 20 µL were used for the gel electrophoresis.
Lane 3: 7 µL of a pKAN- EcoRI/HindIII double digestion. Within the double digestion, one can find 38 µL of pKAN, 3 µL of the EcoRI enzyme, 3 µL of the HindIII enzyme, 6 µL of 10x Buffer 2.1, and 10 µL of water. A total volume of 60 µL was present in our 2 mL tube. Again, only 7 µL were used for the gel electrophoresis.
Lane 4: 12 µL sample of a pAMP purified sample. Within the sample, one can find 5 µL of pAMP, 5 µL of water, and 2 µL of loading dye.
Lane 5: 12 µL sample of a pKAN purified sample. Within the sample, one can find 5 µL of pKAN, 5 µL of water, and 2 µL of loading dye.
Lane 6: 5 µL sample of a pKAMP (pAMP/pKAN)- EcoRI/HindIII double digestion. Within the double digestion, one can find 2 µL of pKAMP, 1 µL of the EcoRI enzyme, 1 µL of the HindIII enzyme, 3 µL of 10x Buffer 2.1, and 23 µL of water. A total volume of 30 µL was present in our 2 mL tube. Again, only 5 µL were used for the gel electrophoresis.
Lane 7: 10 µL of ladder. We began our goal of combining DNA by obtaining the desired samples of plasmid DNA from a liquid bacteria culture. Plasmids are circular pieces of DNA, which can be found in bacteria. Plasmids are often used in the manipulation of genes. After carefully following a mini-prep procedure, we were left with samples of pAMP and pKAN plasmids. pAMP is a bacterial plasmid, which contains the gene ampR. The ampR gene codes for a protein that breaks down the