Nt1310 Unit 1 Lab Report
2. Steps for Southern Blotting
Digest the DNA with an appropriate restriction enzyme.
Run the digest on an agarose gel.
Denature the DNA, which would separate double-stranded DNA into single-stranded DNA.
Transfer the denatured DNA to the membrane.
Probe the membrane.
Visualize your radioactively labeled target sequence. If you used a radiolabeled 32P probe, then you would visualize by autoradiograph.
3. a) She placed the nucleotide as she did because the EcoR I cuts in one place which is why there are two bands. One fragment is slightly smaller than the other.
b) She was able to add this sequence because The Hind III by itself only cut once. The EcoR I also cut once. In the double digest, the Hind III cuts the …show more content…
smaller of the EcoR I fragments.
4.
The odds that the sequence will occur in random order is 1 out of 4096. Since there are four bases possible and they must appear in a six base sequence the answer is 1 out of 4.
5. By digesting the DNA with the above enzyme there would be about then divide 3 billion by 4,096
7. Advantages bacteria gain by having these restriction enzymes is that restriction enzymes serve as a means of defense for a bacterial cell. Restriction enzymes can prevent DNAfrom other organisms, such as bacterial viruses or other bacterial species, from entering the cell and taking over vital cell processes.
9. If you placed the gel with the wells next to the red electrode instead of the black one is that the DNA would still be attracted to the positive electrode. The DNA samples would run quickly toward the top of the gel and out into the buffer.
10. Gel only slightly higher percentage of Agarose would probably not affect the results significantly. However, if the percentage were greatly increased, the larger fragments may not be able to move through the decreased pore size making it difficult, resulting in less effective electrophoretic separation.
11. Agarose gel the substrate used to separate molecules. Contains microscopic pores that allow smaller molecules to move through
faster.
TBE buffer a liquid buffer that protects the DNA molecules and allows electricity to move through the chamber, driving the molecules across the gel.
Electrophoresis chamber holds all of the components of the separation. Creates a closed system in which to submerge a gel and apply electrical current.
Power supply the unit generating the electricity being applied to the chamber.
DNA samples the substance to be separated. A mixture of differently-sized fragments that separate when electricity is applied.
DNA stain used to stain the gel after electrophoresis. Since DNA cannot be seen with the naked eye, DNA stain is necessary to visualize your DNA fragments.
12. Another use for gel electrophotresis is screening for genetic disorders. Genetic screening is a health program that identifies treatable genetic disorders in newborn infants or adults. Early intervention to treat these disorders can eliminate or reduce symptoms that might otherwise cause a lifetime of disability. Benefits using this technology are: our genetic counselor can tell you with greater certainty that you have the same relatively low risk of developing breast cancer as women in the general population. If your test result is abnormal, monitoring can help find cancer at the earliest stage, when it is most treatable and curable. In the case of prenatal testing a diagnosis can be made on a suspect fetus that will allow ample time for decision to be made about treatment. And finally, results from genetic tests can also help predetermine the carrier status of couples planning parenthood.
Digest the DNA with an appropriate restriction enzyme.
Run the digest on an agarose gel.
Denature the DNA, which would separate double-stranded DNA into single-stranded DNA.
Transfer the denatured DNA to the membrane.
Probe the membrane.
Visualize your radioactively labeled target sequence. If you used a radiolabeled 32P probe, then you would visualize by autoradiograph.
3. a) She placed the nucleotide as she did because the EcoR I cuts in one place which is why there are two bands. One fragment is slightly smaller than the other.
b) She was able to add this sequence because The Hind III by itself only cut once. The EcoR I also cut once. In the double digest, the Hind III cuts the …show more content…
smaller of the EcoR I fragments.
4.
The odds that the sequence will occur in random order is 1 out of 4096. Since there are four bases possible and they must appear in a six base sequence the answer is 1 out of 4.
5. By digesting the DNA with the above enzyme there would be about then divide 3 billion by 4,096
7. Advantages bacteria gain by having these restriction enzymes is that restriction enzymes serve as a means of defense for a bacterial cell. Restriction enzymes can prevent DNAfrom other organisms, such as bacterial viruses or other bacterial species, from entering the cell and taking over vital cell processes.
9. If you placed the gel with the wells next to the red electrode instead of the black one is that the DNA would still be attracted to the positive electrode. The DNA samples would run quickly toward the top of the gel and out into the buffer.
10. Gel only slightly higher percentage of Agarose would probably not affect the results significantly. However, if the percentage were greatly increased, the larger fragments may not be able to move through the decreased pore size making it difficult, resulting in less effective electrophoretic separation.
11. Agarose gel the substrate used to separate molecules. Contains microscopic pores that allow smaller molecules to move through
faster.
TBE buffer a liquid buffer that protects the DNA molecules and allows electricity to move through the chamber, driving the molecules across the gel.
Electrophoresis chamber holds all of the components of the separation. Creates a closed system in which to submerge a gel and apply electrical current.
Power supply the unit generating the electricity being applied to the chamber.
DNA samples the substance to be separated. A mixture of differently-sized fragments that separate when electricity is applied.
DNA stain used to stain the gel after electrophoresis. Since DNA cannot be seen with the naked eye, DNA stain is necessary to visualize your DNA fragments.
12. Another use for gel electrophotresis is screening for genetic disorders. Genetic screening is a health program that identifies treatable genetic disorders in newborn infants or adults. Early intervention to treat these disorders can eliminate or reduce symptoms that might otherwise cause a lifetime of disability. Benefits using this technology are: our genetic counselor can tell you with greater certainty that you have the same relatively low risk of developing breast cancer as women in the general population. If your test result is abnormal, monitoring can help find cancer at the earliest stage, when it is most treatable and curable. In the case of prenatal testing a diagnosis can be made on a suspect fetus that will allow ample time for decision to be made about treatment. And finally, results from genetic tests can also help predetermine the carrier status of couples planning parenthood.