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In this activity related to plant biotechnology, learners use restriction enzymes to cut up DNA from a virus called Bacteriophage λ, a process known as restriction digestion. After overnight digestion, the reaction is stopped by addition of a loading buffer. The DNA fragments are separated by electrophoresis, a process that involves application of an electric field to cause the DNA fragments to migrate into an agarose gel. The gel is then stained with a methylene blue stain to visualize the DNA bands and may be photographed. This lab will help learners understand what a DNA restriction enzyme is and how it works, how to use a micropipette, how to separate DNA using electrophoresis, and how to use a restriction digestion map to identify a sample DNA. This lesson guide includes background information, safety precautions and notes, and questions with answers for learners. For safety reasons, adult supervision is recommended.
- 1 to 7 days
- 1 to 7 days
- Over $20 per group of students
- Ages 11 - adult
- Activity, Experiment/Lab Activity, Lesson/Lesson Plan
- English
Quick Guide
Materials List (per group of students)
- Four microtubes
- Microtube rack
- 20-µl micropipette (or 10-µl micropipette) and sterile tips
- Waterproof pen
- Beakers or foam cups with crushed ice
- 20 µl of 0.4 µg/µl λ DNA
- 2.5 µl BamHI restriction enzyme
- 2.5 µl EcoRI restriction enzyme
- 2.5 µl HindIII restriction enzyme
- 10 µl distilled water
- Gloves
- 500-ml beaker
- Electrophoresis chamber
- Power supply
- 20 µl 10X loading dye
- 1.0% agarose gel
- Container with TBE solution
- 37°C water bath w/ floating rack
- 60°C water bath or saucepan on a hot plate
- Cooler with crushed ice
- Freezer (non frost-free, if possible)
- Camera if desired
- Distilled water
- 0.002% methylene blue stain
Subjects
-
Engineering and Technology
-
Engineering
- Bioengineering/Biomedical Engineering
-
Technology
- Agriculture and Biotechnology
-
Engineering
-
Life Sciences
-
Cells
- Cell Structure and Function
-
Diversity of Life
- Viruses and Bacteria
-
Heredity and Genetics
- DNA Structure and Function
- Genetic Engineering
-
Cells
-
Physical Sciences
- Chemistry
-
The Nature of Science
-
The Scientific Process
- Conducting Investigations
- Gathering Data
- Formulating Explanations
- Communicating Results
-
The Scientific Process
Audience
To use this activity, learners need to:
- see
- see color
- read
- touch
Learning styles supported:
- Involves teamwork and communication skills
- Involves hands-on or lab activities
Other
Components that are part of this resource:
- Gel Electrophoresis of Dyes Activity
- Student Activity: Restriction Enzyme Analysis - Methylene Blue stain
This resource is part of:
Access Rights:
- Free access
By:
- Stephens, Janice ; Leach, Jan
Rights:
- All rights reserved, The American Phytopathological Society, 2011