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36 terms

pGLO Transformation Lab

pGlo Lab Study Guide
STUDY
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GFP
Green Fluorescent Protein
Puget Sound of Coastal Washington
Where you can find Jelly Fish with GFP
Aequorea Victoria
Name of Jelly Fish with GFP
238
How many amino acids are in Green Fluorescent protien?
b-can
The structure of green fluorescent protein
beta sheet outside with alpha helis inside
The b-can structure consists of...
the wavelength of the photon
What makes it green?
the protein uses an electron transport chain to produce photons
How does GFP work?
E. coli 101
What strain of bacteria did we use?
nonpathogenic
Term meaning that it is not toxic or harmful to humans
Arabinose
Switches on the GFP gene; must be present for bacteria to glow
Ampicillin
antibiotic that breaks down the cell wall and kills bacteria
ultraviolet light
Provides energy for the luminescence to take place
single celled, reproduces quickly, nonpathogenic
Why E. coli 101 is an optimal choice for a genetic transformation
cream or light yellow
What color are the E. coli colonies?
nutrient agar
What did we grow the bacteria on?
plasmid
small circular pieces of DNA outside of the nucleoid, often exchanged by bacteria in sexual reproduction
transformation solution
CaCl2 (Calcium chloride)
heat shock
The process used to fracture the membrane of the bacteria to insert the plasmid
inoculating loop
Long, thin instrument with a small loop on the end of it, used various times throughout the experiment procedure
no, the protein grows, not the DNA
Did the pGLO DNA solution glow under UV light?
control
Why wasn't pGLO DNA solution inserted into the -pGLO tubes?
LB/amp/ara +pGLO plate
Which plate of bacteria glowed under UV light?
LB/amp +pGLO plate
Which plate of bacteria grew in the prescence of ampicillin?
LB/amp -pGLO plate and LB/amp/ara B
Which plates did not grow any bacteria?
LB -pGLO plate
Which plate grew bacteria most similar to the parent bacteria?
total # of transformed cells/Amount of DNA spread on plate
How to calculate transformation efficiency
9.8125x10^2
981.25 in Scientific Notation
transformation efficiency
A way to determine how effective the transformation was
promoter, operater, ampicillin resistence, GFP
Genes on the genetically engineered plasmid inserted into the E. coli
add our own operon to transmutate the bacteria
Goal of this experiment
how bacteria sexually reproduce
This experiment demonstrated
stiffened the membrane
Putting the tubes on ice initially
ractured the membrane
Moving the tubes from ice to the water bath
kept it fractured
Moving the tubes from the water bath back to the ice
Their waste
what about bacteria makes you sick?