What is the purpose of the pGLO transformation lab?
pGLO Bacterial Transformation and GFP Kits pGLO Lab Kits utilize Bio-Rad’s pGLO plasmid, which encodes a green fluorescent protein (GFP), to enable instructors to give students a hands-on introduction to transformation, cloning, protein chromatography, and electrophoresis techniques.
What was the purpose of the heat shock step in the pGLO lab?
Plasmid DNA is added to half of the cells before they are “heat shocked” in a 42°C water bath. The heat shock step facilitates the entry of DNA into the bacterial cells. Recovery Broth is added to the cell suspension, and the bacteria are allowed to recover for 30 minutes at 37°C.
What are the three important genes located on the pGLO plasmid?
The pGLO plasmid contains an origin or replication, a selectable marker, and the gene for Green Fluorescent Protein (GFP). The plasmid also contains a gene for the arabinose C protein, which is a protein that regulates expression from the arabinose BAD promoter (PBAD).
What two factors must be present in the bacteria’s experimental environment for you to see the green color?
What two factors must be present in the bacteria’s environment for you to see the green color? (Hint: one factor is in the plate and the other factor is in how you look at the bacteria). The sugar arabinose in the agarose plate is needed to turn on the expression of the GFP gene.
What factors influence transformation efficiency?
The factors that affect transformation efficiency are the strain of bacteria, the bacterial colony’s phase of growth, the composition of the transformation mixture, and the size and state of the foreign DNA.
Why can the bacteria cells that have been genetically transformed grow on LB AMP plates?
If there are any genetically transformed bacterial cells they would be located on the +pGLO LB/amp and the +pGLO LB/amp/ara plates. This is because both these plates have the plasmid inside them which allows for the transformation.
How does temperature affect transformation efficiency?
Previous experiments have demonstrated increased electroporation transformation efficiencies for cells grown at lower temperatures (3); therefore, we hypothesized that cells grown at the lower temperature of 20°C will have a higher transformation efficiency as compared to cells grown at 37°C.
Why does pGLO glow under UV light?
Because it shares a bidirectional promoter with a gene for metabolizing arabinose, the GFP gene is expressed in the presence of arabinose, which makes the transgenic organism express its fluorescence under UV light.
Why was ampicillin added to some of the plates in the transformation lab?
This plasmid contains an ampicillin-resistance gene in addition to the GFP gene. Ampicillin is an antibiotic and works by preventing E. coli from constructing cell walls, thereby killing the bacteria.
How would you change the bacteria’s environment the plate they are growing on to best tell if they are ampicillin resistant explain?
2. How would you change the bacteria’s environment to best tell if they are ampicillin resistant? The best test would be to take some of the bacteria growing on the LB plate and streak them on an LB/amp plate. If the bacteria are viable on the LB/amp plate, then they are resistant to ampicillin.
Which plate would be most likely to indicate transformed cells?
Which plate would be most likely to indicate transformed cells? A plate with 100 colonies growing on LB agar with kanamycin.
How many terms are in the pGLO transformation lab?
pGLO Lab Analysis 16 terms gracebronner Bacterial Transformation Lab: pGLO 34 terms mocha_dog529 Bio Lab Midterm 88 terms aya2 Sets with similar terms pGLO Transformation 19 terms Ayah_Rasheed Biomed Unit 7 57 terms Angela_Silva32 Bacterial Transformation Lab 16 terms GermaineRicablanca Microbiology Lab Exam 115 terms BrittB95
What is PGLO and how does it work?
A description of what you’ll be working with, including a brief overview of transformation. pGLO: How the plasmid works. In this lab, you’ll transform E. coli cells with the plasmid pGLO, which contains a gene for green fluorescent protein (GFP). This will allow you to grow bacterial colonies that are bright, fluorescent green.
How much does pGLO lab cost?
Subjects Explanations Create Study sets, textbooks, questions Log in Sign up Upgrade to remove ads Only $35.99/year pGlo Lab Questions – AP Biology STUDY Flashcards Learn Write Spell Test PLAY Match Gravity Created by keelin_vic
How many terms are in the pGLO AP Biology test?
pGlo Lab Test Review – AP Biology 28 terms keelin_vic pGLO Lab Analysis 16 terms gracebronner Bacterial Transformation Lab: pGLO 34 terms mocha_dog529 Bio Lab Midterm 88 terms aya2 Sets with similar terms pGLO Transformation 19 terms Ayah_Rasheed Biomed Unit 7 57 terms Angela_Silva32 Bacterial Transformation Lab 16 terms GermaineRicablanca