Secrets of the Rainforest™ Kit

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Secrets of the Rainforest Kit

Overview

Lab Preparation Checklist
Kit contains sufficient materials for 8 student workstations. (2–4 students per workstation)
Module 1: Preparing a Bacterial Library  
Bacterial library, E. coli, lyophilized 1
LB nutrient agar tablets 5
Petri dishes, 60 mm, sterile 20
Inoculation loops, sterile 20
Disposable plastic transfer pipets 10
Module 2: Protein Purification  
Ampicillin, lyophilized 1
Arabinose, lyophilized 1
Lysozyme, lyophilized 1
LB nutrient broth tablet 1
Binding buffer 1
Column equilibration buffer 1
Column wash buffer 1
Elution buffer 1
Chromatography columns 8
Inoculation loops, sterile 20
Disposable plastic transfer pipets 40
Micro test tubes, 2.0 ml, clear 30
Cell culture tubes, 15 ml, sterile 25
Sample collection tubes, 5 ml 25
Foam micro test tube holders 8
Curriculum, including Teacher's Guide, Student Manual, and graphic Quick Guide 1
Required Accessories Not Included in Kit:
UV lamps 4–8
Centrifuges 1–2
Recommended (Optional) Accessories:
Incubation oven  
Rocking platform  
Microwave oven  
Secrets of the Rainforest Kit Flowchart
Secrets of the Rainforest Kit Flowchart
Key Kit Features
  • Grow a bacterial library
  • Clone a gene of interest in bacteria
  • Express a protein of interest in bacteria
  • Purify a protein of interest from bacteria
  • Take a protein to market
  • Complete in four 45 min lab sessions
Curriculum Fit
Curriculum Fit
Creating a Bacterial Library and Cloning a Gene

How is a bacterial library created and a gene cloned? A bacterial library is constructed by chopping up the genome of the medicinal plant (the "parent") with a restriction enzyme. Restriction enzymes recognize specific double-stranded DNA sequences and cut each DNA strand, often creating "sticky ends" known as 3' or 5' overhangs. This generates a random assortment of DNA fragments. Some of these DNA fragments from the medicinal plant will contain the gene that encodes the green glowing protein with anticancer properties. When the same restriction enzyme is used to cut (digest) a plasmid (the "destination") it produces an opening in the plasmid with sticky ends that are complementary to the sticky ends of the medicinal plant DNA fragments. Next, the plant DNA fragments are inserted into plasmids by joining (ligating) their sticky ends and covalently bound together by a DNA ligase enzyme. The resulting plasmids contain random assortments of DNA fragments derived from the genome of the medicinal plant. The plasmids are transformed into the bacteria, E. coli. When transformed bacteria divide, the plasmid is also replicated. In the best-case scenario, some of the transformed bacterial cells will contain copies of the plasmid containing the plant gene, which encodes the protein of interest.

Secrets of the Rainforest colonies and GFP separation on a series of hydrophobic interaction chromatography (HIC) columns.

Secrets of the Rainforest colonies and GFP separation on a series of hydrophobic interaction chromatography (HIC) columns.

The Secrets of the Rainforest™ Kit provides a biotechnology curriculum that takes students on an adventure starting in the rainforest in the Andes, continuing to a biotechnology company engaged in developing new pharmaceutical compounds, and finally to the Food and Drug Administration.

Features and Benefits

  • Applies the principles of DNA restriction analysis, bacterial transformation, and protein purification
  • Introduces your students to the world of commercial biotechnology

With this kit, students are able to:

  • Grow a bacterial library
  • Clone a gene of interest in bacteria
  • Express a protein of interest in bacteria
  • Purify a protein of interest from bacteria
  • Simulate taking a protein to market

How It Works

In the hands-on portion of the exercise, students streak out the bacterial library, grow the bacteria, and select the colonies that have acquired the green glowing trait from a background of nonglowing bacteria. A culture of bacteria is grown to scale up production of the green glowing protein, which is then purified by column chromatography. The actual source of this fluorescent protein is the bioluminescent jellyfish Aequorea victoria.

This practical activity is followed by dry-lab lessons in which the purified green protein is put through the drug discovery process. This includes learning about Food and Drug Administration regulations, animal testing, marketing practices for introducing a new drug, financing, ethical issues, and student presentations. Students examine the needs and viewpoints of advocacy groups, the biotechnology industry, and the FDA. They consider ethical, economic, social, and practical issues, and develop possible strategies for what they feel are the best outcomes.

More Information

  • Four-session laboratory activity, 45 min per session
  • Provides sufficient materials for eight student workstations, up to four students per workstation
Kit contains sufficient materials for 8 student workstations (2–4 students per workstation)  
Module 1: Preparing a Bacterial Library  
  Bacterial library, E. coli, lyophilized 1
  LB nutrient agar tablets 5
Petri dishes, 60 mm, sterile 20
Inoculation loops, sterile 20
Disposable plastic transfer pipets 10
Module 2: Protein Purification  
Ampicillin, lyophilized 1
Arabinose, lyophilized 1
Lysozyme, lyophilized 1
LB nutrient broth tablet 1
Binding buffer 1
Column equilibration buffer 1
Column wash buffer 1
Elution buffer 1
  Chromatography columns 8
Inoculation loops, sterile 20
Disposable plastic transfer pipets 40
Micro test tubes, 2.0 ml, clear 30
Cell culture tubes, 15 ml, sterile 25
Sample collection tubes, 5 ml 25
Foam micro test tube holders 8
Curriculum, including Teacher's Guide, Student Manual, and graphic Quick Guide 1
Secrets of the Rainforest Kit

166-0006EDU
Bacterial library screening kit, classroom study kit, includes bacterial library and protein purification modules, curriculum, and more, for 32 students; education use only

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3051 Secrets of the Rain Forest Kit, Biotechnology Explorer, Quick Guide Click to download
4006098 Instruction Manual, Secrets of the Rain Forest Kit, Biotechnology Explorer, Rev B Please login to download this document [ Add to Cart (Free) ]