This is the experimental plate. Why do only transformants grow on this plate? Make sure to also include your lab group name and the date. Which plates are the control plates?
Clean the lab benches with the bleach solution and remember to wash your hands before leaving the lab. Because of the simple structures of bacterial cells, they are the most commonly modified organisms. The final recombinant model plasmid, a merger of the original plasmid DNA and the added insulin gene.
Genes are removed using restriction enzymes. Will all of the plates have bacteria growing on them? Many times, and in this activity, a gene is simply added to the bacteria, causing the bacteria to be able to produce a useful protein, such as insulin.
In blue, the model plasmid after being cut by a restriction enzyme. Bacteria are so plentiful that a ounce bottle of water may contain up to million bacteria! The protocol above has been modified from UC Davis. First, a desired gene must be selected from some any organism, that is, the gene that codes for the creation of insulin protein, and removed from the DNA of that organism.
Make sure that you add to the "-" tube first so as to avoid cross-contamination of the plasmid. Tape the ends of the gene to the matching sticky end on the plasmid see Figure 5. Micrometers are one millionth of a meter.
Estimates vary from 10 million to 1 billion species. Explain your answer to question 2. These enzymes search for specific nucleotide sequences in the DNA, called recognition sites, where they "cut" the DNA by breaking certain bonds.
Now observe the results on the petri dishes without removing the lids. Yes Bacteria are the most commonly modified organisms. Bacteria are everywhere, and most of the time they are harmless.
Place in a rack and incubate for 10 minutes at room temperature. Have them staple their recombinant models on their worksheets and complete the worksheets as a team.
If the DNA sequences have been correctly cut, the base pairs from each end of the gene match exactly with the cuts made in the plasmid. On plate 4 any bacteria that has been transformed by taking up the pGREEN plasmid is now able to grow in the presence of the antibiotic since the plasmid also contains the gene allowing for antibiotic resistance.
Let them stay on ice for at least one minute.
Conclude the activity by administering the Assessment Questions, as described in the Assessment section. Complete the remainder of the activity sheet you began on day 1 of the lab. Wash hands before leaving lab. National Institutes of Health http: In order to determine the efficiency of the transformation we need to determine the initial amount mass of plasmid that was spread on the plate and relate this to the number of transformed colonies that were observed on the experimental plate.
When the bonds are broken in a staggered manner it creates "sticky ends.Hands-on Activity: Bacteria Transformation Contributed by: National Science Foundation GK and Research Experience for Teachers (RET) Programs, University of Houston they isolate a mammal insulin gene and combine it with a bacteria's gene sequence (plasmid DNA) for production of the protein insulin.
Modeling Bacteria Transformation. BACTERIA TRANSFORMATION LAB (ACTIVITY) In this exercise you will use paper to simulate the cloning of a gene from one organism into a bacterial plasmid using a restriction enzyme digest.
Transforming E. Coli with pGLO Plasmids, a Lab As a class draw and take notes on the diagram that depicts how the genes in the plasmid interact with each other The Lab Objective: 1. With the tools and lab protocol provided, you will be able to perform genetic transformation. By the end of the lab activity and analysis you will.
Activity 4: Transformation of E.
coli using green Wash hands thoroughly before leaving lab. Student Activity: Transformation of the bacterium E. coli using a gene discrete spots around the bell margin of the jellyfish and will fluoresce under certain conditions When inserted into a plasmid and used for the transformation procedure.
LabBench Activity Key Concepts I: Bacterial Transformation. Genetic transformation occurs when a host organism takes in foreign DNA and expresses the foreign gene. In this part of the lab, you will introduce a gene for resistance to the antibiotic ampicillin into a bacterial strain that is killed by ampicillin.
Biology Laboratory Manual – page 96 Activity #5b. Plasmid DNA Isolation, Restriction Enzyme Digestion & Bacterial Transformation Learning Goals.Download