| 
  • If you are citizen of an European Union member nation, you may not use this service unless you are at least 16 years old.

  • Social distancing? Try a better way to work remotely on your online files. Dokkio, a new product from PBworks, can help your team find, organize, and collaborate on your Drive, Gmail, Dropbox, Box, and Slack files. Sign up for free.

View
 

Lesson 4-5: Transformation Wet Lab

Page history last edited by mariaelizabethbunn@... 4 years, 7 months ago
Time

Engaging the Student (Entry Task) 

Developing the Ideas--Lesson

Checking for Understanding (exit ticket)

Student Handout 
Teacher/Lesson Notes
Materials

~80-100 minutes**

 

**This lab requires a great deal of preparation in advance. You will need to make and pour two different types of agar plates which should be poured a couple days before the lab. Preparing plates for one class will take a minimum of 2-hours.

 

If you have 50-minute class periods, there is a stopping point in the transformation procedure after the heat-shock step. It will take students between 40-50 minutes to get to this point. Bacteria can be refrigerated until the next day when you're ready to continue. Refer to the Teaching Tips provided. 

 Show students some of the glowing animals pictured at the National Geographic website, here

 

Ask students:

 

  • How did scientists made these animals glow?
  • How does the technique they used relate to the creation of human insulin in the lab?

 

Introduce the transformation lab and explain what students will be doing. The PBLU plasmid map link (see Teacher/Lesson notes) may be a useful visual.  

View the full lesson plan here:

Supplemental teaching resources:

 

In this wet lab, students will work on a procedure for inserting the pBLU© plasmid DNA into E. coli bacteria. If the transformation is successful, students will see that the E.coli bacteria now have a blue color that is visible under full spectrum light (sun/artificial room lighting), and fluoresce upon illumination with a black light. In the context of the Diabetes project, the pBLU© plasmid will represent the human insulin gene (INS). Teachers have the option to let students work on optimizing the transformation procedure as a second component to the lab.

 

If you are a member of SEP (or know MaryMargaret Welch), you may have access to borrow the kit for this lab provided by Fred Hutch. The information and documents provided in this lesson plan are taken from the SEP kit. Note that the kit provides everything except: a heat source to melt the agar (microwave or hotplate), ethanol or bleach, and ice. These must be provided by the teacher. 

 

If you do not have access to the SEP kit, you may buy the "pGLO Bacterial Transformation Kit" from Bio-Rad. Full information on purchasing and kit inventory can be found here

Note: This materials list is comprehensive only if using the Fred Hutch SEP kit (see Teacher/Lesson Notes); if purchasing a kit from Bio-Rad you will need to assess its inventory and determine any additional materials needed.

 

  • Microwave or hotplate/stovetop
  • 70% Ethanol or 10% bleach solutions
  • Ice 
  1. What evidence from you lab indicates that he bacteria became resistant to ampicillin? (or did not become resistant).
  2. What are three factors that might influence the success of this transformation procedure?
  3. Why would it be advantageous for a bacterium to have an antibiotic resistant gene in its DNA? Why might that be bad for us? 

 

 

Comments (0)

You don't have permission to comment on this page.