Virtual Laboratory Bacterial Transformation Name: ________________________________ Introduction 1. Log onto the Pearson LabBench web page at http://www.phschool.com/science/biology_place/labbench/index.html 2. Select Lab 6 – Molecular Biology / Part I – Transformation in Bacteria 3. Follow the instructions carefully on each screen – be sure to review all animations. 4. Answer the lab packet questions as you proceed through the virtual lab. 1. Read “Key Concepts I: Bacterial Transformation”. What is bacterial transformation? _______________________________________________________________________________________ _______________________________________________________________________________________ a. What gene are we going to attempt to introduce into our bacterial strain? _________________________________________________________________________________ b. What will happen to our bacteria strain if we are successful? _________________________________________________________________________________ 2. Read “Bacterial Colonies”. What species of bacterial are we using for this experiment? _______________________________________________________________________________________ 3. Read “E. coli Bacteria”. Where is E. coli bacteria commonly found? _______________________________________________________________________________________ _______________________________________________________________________________________ a. How does E. coli reproduce? How rapidly can this occur? _________________________________________________________________________________ _________________________________________________________________________________ b. Describe the chromosomal genome of E. coli. _________________________________________________________________________________ _________________________________________________________________________________ _________________________________________________________________________________ 4. Read “Plasmids”. What are plasmids? _______________________________________________________________________________________ _______________________________________________________________________________________ a. How are plasmids used in genetic engineering? _________________________________________________________________________________ _________________________________________________________________________________ L. Carnes Adapted from Pearson: LabBench Activities MVHS AP Biology http://www.phschool.com/science/biology_place/labbench/index.html b. What does the ampR gene confer resistance to in E. coli? _________________________________________________________________________________ _________________________________________________________________________________ c. What happens to E. coli cells that do not carry the ampR gene? _________________________________________________________________________________ _________________________________________________________________________________ d. In order to transform cells...what do we need to do first? _________________________________________________________________________________ _________________________________________________________________________________ 5. Read “Competent Cells”. Describe what this concept means. _______________________________________________________________________________________ _______________________________________________________________________________________ _______________________________________________________________________________________ _______________________________________________________________________________________ a. What processes are used to make cells competent? _________________________________________________________________________________ _________________________________________________________________________________ b. In what phase of bacterial growth is it easiest to make cells competent? _________________________________________________________________________________ _________________________________________________________________________________ 6. Read “Design of Experiment I”. Make notes of some important safety procedures: _______________________________________________________________________________________ _______________________________________________________________________________________ _______________________________________________________________________________________ _______________________________________________________________________________________ 7. Read “Transformation Procedure”. After you've familiarized yourself with the procedure as a whole, take a closer look at each stage. Select steps 1–4 and 6 to see what is going on at the cellular level. THE VERY LAST “A Closer Look” screen shows an animation of the transformation procedure...WATCH IT! There are basically SIX steps to follow to correctly transform bacteria. Describe these steps IN ORDER below: a. _________________________________________________________________________________ _________________________________________________________________________________ b. _________________________________________________________________________________ _________________________________________________________________________________ c. _________________________________________________________________________________ _________________________________________________________________________________ d. _________________________________________________________________________________ _________________________________________________________________________________ e. _________________________________________________________________________________ _________________________________________________________________________________ f. _________________________________________________________________________________ _________________________________________________________________________________ 8. The figures below show the events that take place during transformation of an E. coli cell. Type in the letters to indicate the correct order in which the events occur. 9. Read “Analysis of Results I”. Draw AND describe the four possible outcomes for this experiment in the space provided below. a. What does it mean if there is a “lawn growth of cells? _________________________________________________________________________________ _________________________________________________________________________________ b. What does it mean if there are individual colonies of growth on the agar? _________________________________________________________________________________ _________________________________________________________________________________ c. What does it mean if nothing is growing on the agar? _________________________________________________________________________________ _________________________________________________________________________________ 10. Read “Label the Results of Your Experiment”. After incubation, the following plates were removed from the incubator. All but part of one label have been removed so that you must now use your understanding of this laboratory to make new labels for each plate. Based on the PHYSICAL APPEARANCE of each plate, select the appropriate label for the plates. DESCRIBE THIS BELOW...do NOT simply write A, B, C, or D! For example...Plate I should be labeled ______________________ because _________________________. a. Plate I: _________________________________________________________________________________ _________________________________________________________________________________ b. Plate II: _________________________________________________________________________________ _________________________________________________________________________________ c. Plate III: _________________________________________________________________________________ _________________________________________________________________________________ d. Plate IV: _________________________________________________________________________________ _________________________________________________________________________________ 11. Take “Lab Quiz I”. You may write your responses below: □ Question #1 _____ □ Question #2 _____ □ Question #3 _____ □ Question #4 _____ □ Question #5 _____ □ Question #6 _____