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pGlo Lab Study Guide

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?

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