Some bacterial species have been found to take in DNA from the environment, and the newly acquired DNA may give them a selective advantage. Scientists can take advantage of this phenomenon by purposely transforming bacteria or inducing them to take up DNA. This method is used to study gene function and gene expression in bacteria and can have clinical relevance when bacteria take up an antibiotic resistance gene. Clinicians routinely take samples from patients, and if a microorganism is present in a patient sample, its genome is also present. To detect and identify the microorganism, scientists can assay for the presence of an organism's DNA using a powerful molecular biology and diagnostic technique known as the polymerase chain reaction (PCR). The PCR products can be analyzed using agarose DNA gel electrophoresis. Students will complete the midterm exam at the end of this topic.
Objectives:
- Describe the method of bacterial transformation with plasmid DNA.
- List the DNA sequences/genes present on the pGLO plasmid and explain the function of each.
- Illustrate and interpret the phenotypic traits of transformed bacteria observed on non-selective and selective media.
- List the components needed to set up a polymerase chain reaction (PCR) reaction.
- Describe the three steps in a PCR cycle illustrating what happens in each step using DNA, primers, and Taq DNA polymerase.
- Explain how PCR cycles create copies of target DNA.
- Explain the theory of agarose DNA gel electrophoresis, analyze a PCR reaction using agarose DNA gel electrophoresis, and illustrate and describe the results.
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