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pGLO Transformation Lab Quiz
Terms in this set (34)
What is the procedure called that you will be overall preforming in this lab?
piece of DNA that provides the codes for making a protein
What does the protein do that is made from the genes that are coded?
it gives an organism a particular trait
change caused by genes
green fluorescent protein
What is the scientific name for the jellyfish that exemplifies the GFP?
If the procedure goes correctly what should happen?
it should express the gene by glowing an ultraviolet green color as it produces the GFP
What should be learned from this lab?
the process of moving genes from one organism to another with the aid of a plasmid
In addition to one large chromosome, what else is bacteria known for having?
small circular pieces that hold plasmid DNA
What is special about what plasmid DNA carries?
they carrie the genetic code for proteins that are beneficial to bacterial surfaces
What natural mechanism allows bacteria to adapt to new environments?
bacteria transferring plasmids back and forth allowing them to share the beneficial genes
What occurs due to the transmission of plasmids?
bacterial resistance to antibiotics
What does Bio-Rad's pGLO plasmid encode for?
the GFP gene and another gene fir antibiotic ampicillin
incorporates a special gene in the regulatory system, which can be used to control expression of the fluorescent protein in transformed cells
How can the GFP gene be switched on in transformed cells?
by adding sugar arabinose to the cells' nutrient medium
How do selection for cells that have been transformed with pGLO DNA?
growth on ampicillin plates
What color should transformed cells appear for the plates that do NOT contain arabinose?
white (wild-type phenotype)
What color should transformed cells appear for the plates that do contain arabinose under a UV light?
What is the first step in the lab?
label one tube -pGLo and one +pGLO
What is the second step in the lab?
put 250ml of transformation solution in each tube
What is the third step in the lab?
place the tubes on ice
What do use to pick up colonies of bacteria in step 4?
a sterile loop
What else happens in step 4?
-select starter colonies
-choose colonies that are circular and smooth edges
-spin the loop between your index and thumb fingers until the whole colony is dispersed with no floating chunks
-place the tube back in the ice
What happens in step 5?
-examine pGLO DNA with UV lamp
-get a new sterile loop
-there should be a film of plasmids that look like soapy bubbles
-mix lapful into pGLO tube
-you should have a -pGLO and a +pGLO tube
What is the sixth step in the lab?
incubate the tubes (so that the bottoms are touching the ice) on ice for 10 minutes
What is step 7?
label the 4 LB nutrient agar plates:
What are the two +pGLO plates in step 7?
LB and ampicillin but one also has arabinose
What are the two -pGLO plates in step 7?
LB but one can ampicillin
What do you do in step 8?
a heat shock
What do you do in step 9?
-remove ice rack
-add 250 ml to the tubes
-incubate for 10 minutes at room temp
What do you do in step 10?
-gently flicked the closed tubs with your finger and resuspend the bacteria
-put suspensions onto the appropriate agar plates
What do you do in step 11?
use a new sterile loop for each plate
-flat the agar but don't press too deep
What do you do in step 12?
-stack the 4 plates and tape them together
-place them upside down in a 37 degree C incubator.
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