What is the purpose of a pBLU lab?
The purpose of this lab was to grow colonies of E. coli.
What is pBLU plasmid?
pBLU is a commercially produced bacterial plasmid that contains genes for ampicillin resistance (beta lactamase and beta galactosidase). It is often used in conjunction with an ampicillin-susceptible E. coli strain to teach students about transformation of eubacteria. It is 5,437 base pairs long.
What was the importance of the LB amp plate in this exercise?
The -DNA/LB plate shows us approximately how many total cells were present in the same volume. This would be only an estimate, as the latter plate would most likely contain a lawn, where you’d have to estimate the number of possible cells.
Why is ampicillin added to the LB medium?
Preparation of Ampicillin Ampicillin is an antibiotic used to selectively eliminate bacteria that have not been transformed with plasmids containing an ampicillin resistance gene.
Why would you want to transform cells?
The phenomenon of transformation has been widely used in molecular biology. As they are easily grown in large numbers, transformed bacteria may be used as host cells for the following: to make multiple copies of the DNA. in cloning procedures.
Where is pGLO from?
The pGLO plasmid is an engineered plasmid used in biotechnology as a vector for creating genetically modified organisms. The plasmid contains several reporter genes, most notably the green fluorescent protein (GFP) and the ampicillin resistance gene. GFP was isolated from the jelly fish Aequorea victoria.
What is the purpose of the LB amp plate (+) plasmid?
The purpose is to successfully transform the Escherichia coli bacteria when we expose it to extracellular plasmid DNA that contains the pGreen gene and the gene for ampicillin resistance.
What are the two genes on the PBLU plasmid?
These reporter genes are very useful in many cloning procedures that will be described in class. Additional information on these two marker genes can be found on the last page of the lab. The plasmid used in this lab that contains these two genes is called PBLUⓇ, BamHI 3160 ECORI 3139 Hindill 3190 4.
What is LB and amp?
LB Amp is Lysogeny Broth (LB) containing the antibiotic ampicillin. Because LB is a rich medium for growing bacteria, adding ampicillin provides a means of selecting transformants that have taken up plasmid DNA containing the bla gene, which encodes resistance to ampicillin.
Why is there no growth on LB ampicillin?
The bacterium cannot grow in the presence of the antibiotic ampicillin unless it contains the plasmid, and so there will be no growth on the LB/Amp plate of the bacteria without the plasmid.
What is the significance of transformation?
The main reason order is significant is that transformations like rotation and scaling are done with respect to the origin of the coordinate system. Scaling an object that is centered at the origin produces a different result than scaling an object that has been moved away from the origin.