Description
Product Details
Advanced—For experienced high school and college classes; requires some technical skill.
Bring your biotechnology classroom instruction to the next level with a unique synthetic biology lab that focuses on the regulation of transcription instead of a traditional exploration of transcription. Students get hands-on experience with cloning ribosomal binding sites onto a plasmid and then observe its effects on transformed bacteria compared to a control by quantifying the expression of GFP in red and/or blue colonies. A truly one-of-a-kind and colorful lab experience that makes it easy to observe the regulation of translation while providing students exposure to a wide breadth of techniques and concepts.
In this lab, students use a thermocycler and the Golden Gate Assembly (GGA) technique to clone 2 different ribosome binding sites (RBS)—a weak ribosome binding site and a strong ribosome binding site—into a plasmid with a reporter gene. They then transform their GGA reactions into E. coli and observe and quantitate the ability of each cloned RBS to support expression of a reporter gene. Students quantitate reporter gene expression using photographs of their plates with the free and easy-to-use ImageJ software.
Product Features
- Designed for 8 groups of 2 to 4 students
- Teacher guide includes paper model activity to help visualize molecular processes
- Includes voucher to request perishable materials at a later date
- Comes with FREE 1-year access to digital resources that support 3-dimensional instruction for NGSS
Time Requirement
Lab, minimum of five 50-minute class periods (250 minutes total). Additional 110 minutes to produce representations of data and discuss outcomes at conclusion of lab.
Digital Resources
Includes 1-year access to digital resources that support 3-dimensional instruction for NGSS. Digital resources may include a teacher’s manual and student guide, pre-lab activities and setup videos, phenomenon videos, simulations, and post-lab analysis and assessments.
Performance Expectation(s)
HS-LS1-1: From Molecules to Organisms: Structures and Processes
Crosscutting Concepts
Cause and Effect
Scale, Proportion, and Quantity
Disciplinary Core Ideas
LS1.A: Structure and Function
LS3.A: Inheritance of Traits
Science and Engineering Practices
Analyzing and Interpreting Data
Engaging in Argument from Evidence
Learning Objectives
- Students learn about gene regulation and can describe the role of ribosome binding sites in producing the appropriate amount of each protein at the correct time and in the appropriate cells.
- Students clone different ribosome binding sites into a plasmid with reporter genes for measuring how well each ribosome binding site supports translation.
- Students explain the difference between type II and type IIS restriction enzymes as well as the significance of the type IIS restriction enzyme cutting pattern in Golden Gate Assembly.
- Students can explain how the cloning technique Golden Gate Assembly works.
- Students transform a complete GGA reaction into E. coli and plate the transformation onto selective media.
- Students make a claim about how well different cloned ribosome binding sites support translation by observing the color and color intensity of the colonies on the plates.
- Students support their claim about how well the different ribosome binding sites support translation by measuring the levels of the reporter protein in E. coli colonies.
Prerequisite Knowledge and Skills
Students should possess a firm understanding of basic DNA structure, a solid understanding of genes and their function, and a solid understanding of the fact that genes are transcribed into RNA that is then translated to form proteins. They also should have a basic understanding of gene expression and regulation, including an understanding of promoters as well as a basic understanding of translation. Experience using a micropipet is needed.