About This Chapter
Basic Molecular Biology Laboratory Techniques - Chapter Summary and Learning Objectives
The lessons in this chapter begin with a definition of genetic engineering. You then can learn about DNA plasmid and its relation to genetic engineering. You'll also study more advanced topics like the functions of restriction enzymes and ligase. Lessons cover the process of agarose gel electrophoresis and how to analyze its results. You'll also be introduced to the processes of bacterial transformation and polymerase chain reactions. The chapter closes with an examination of the Sanger method of DNA sequencing. After completing this chapter, you should be able to:
- Understand genetic engineering
- Identify the function of restriction enzymes
- Explain the procedure of agarose gel electrophoresis
- Define the process of bacterial transformation
- Name the steps and products used in a polymerase chain reaction
- Analyze the Sanger method of DNA sequencing
|What Is Genetic Engineering? - Definition and Examples||Provide a definition and multiple examples of genetic engineering.|
|What Is a DNA Plasmid? - Importance to Genetic Engineering||Explain the importance of DNA plasmids to the process of genetic engineering.|
|Restriction Enzymes: Function and Definition||Recognize the function of restriction enzymes and how they relate to molecular biology.|
|How Ligase Is Used to Engineer Recombinant DNA||Understand how to use ligase to engineer recombinant DNA.|
|What Is Agarose Gel Electrophoresis?||Give a definition of agarose gel electrophoresis.|
|Ethidium Bromide, Loading Buffer and DNA Ladder: Visualizing DNA and Determining its Size||Explain how DNA can be visualized and quantified.|
|Agarose Gel Electrophoresis: Equipment and Procedure||Explain the procedure and equipment needed for agarose gel electrophoresis.|
|Agarose Gel Electrophoresis: Results Analysis||Analyze the results of agarose gel electrophoresis.|
|Bacterial Transformation: Definition, Process and Genetic Engineering of E. coli||Understand bacterial transformation and how it relates to the genetic engineering of E. coli.|
|Bacterial Transformation: Antibiotic Selection and Positive and Negative Controls||Recognize the positive and negative controls used in antibiotic selection.|
|PCR: Reagents Used in Polymerase Chain Reaction||Name the reagents used in polymerase chain reactions.|
|PCR: Steps Involved in Polymerase Chain Reaction||List the steps needed for polymerase chain reactions.|
|The Sanger Method of DNA Sequencing||Define the Sanger method of DNA sequencing.|
1. What is Genetic Engineering? - Definition and Examples
How do we make the insulin used by diabetic patients? In this lesson, you'll learn the basics of how genetic engineering can be used to transform a bacterial host cell into a genetically-modified organism that produces human insulin.
2. What is a DNA Plasmid? - Importance to Genetic Engineering
DNA plasmids play an integral part in most genetic engineering experiments. In this lesson, you'll learn about key features of a plasmid, such as a multiple cloning site, an origin of replication, and a selectable marker.
3. Restriction Enzymes: Function and Definition
Restriction enzymes played a critical role in the advent of genetic engineering. In this lesson, you will learn what role restriction enzymes play in creating recombinant DNA.
4. How Ligase is Used to Engineer Recombinant DNA
DNA ligase makes recombinant DNA technology possible. In this lesson, you will learn how new versions of genes can be designed for experiments in novel host organisms using DNA ligase.
5. What is Agarose Gel Electrophoresis?
Agarose gel electrophoresis plays a key role in genetic engineering experiments. In this lesson, you'll learn what agarose is and how electrophoresis works. You'll also discover the use of agarose in this procedure.
6. Ethidium Bromide, Loading Buffer & DNA Ladder: Visualizing DNA and Determining its Size
In this lesson, you will learn about the role that ethidium bromide, loading buffers, and DNA ladders play in visualizing DNA and determining the size of DNA fragments in agarose gel electrophoresis.
7. Agarose Gel Electrophoresis: Equipment & Procedure
This lesson will review the concepts and mechanisms of agarose gel electrophoresis. It will also summarize the equipment needed to perform the procedure for DNA analysis.
8. Agarose Gel Electrophoresis: Results Analysis
Gel electrophoresis is used to analyze DNA restriction digest and ligation experiments. In this lesson, you will learn how to use a DNA ladder to interpret experimental results.
9. Bacterial Transformation: Definition, Process and Genetic Engineering of E. coli
How can a plasmid be inserted into a bacterial cell? How can transformed bacteria carrying a recombinant plasmid be distinguished from untransformed counterparts? These questions and more will be answered in this lesson.
10. Bacterial Transformation: Antibiotic Selection and Positive & Negative Controls
The use of antibiotic selection and positive and negative controls are important elements of interpreting data from a bacterial transformation. In this lesson, you will learn how antibiotic selection results in colony formation and how controls help pinpoint the cause of experimental problems.
11. PCR: Reagents Used in Polymerase Chain Reaction
Polymerase chain reaction (PCR) is a biotechnology technique that is used to amplify pieces of DNA. In this lesson, you will learn about five ingredients necessary to perform PCR: template DNA, nucleotides, primers, buffer and Taq polymerase.
12. PCR: Steps Involved in Polymerase Chain Reaction
In this lesson, you will learn about the steps required to amplify DNA during PCR. The lesson will explain the role template DNA, primers, nucleotides, Taq polymerase and PCR buffer play in the denaturation, annealing and extension steps of PCR.
13. The Sanger Method of DNA Sequencing
The ability to determine the DNA sequence of an individual is a powerful tool for paternity questions and criminal investigations, among other uses. This lesson will describe one laboratory method that can be used to sequence DNA.
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