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Negative selection through replica plating to screen for ampicillin sensitive colonies. Replica plating is a microbiological technique in which one or more secondary Petri plates containing different solid (agar-based) selective growth media (lacking nutrients or containing chemical growth inhibitors such as antibiotics) are inoculated with the same colonies of microorganisms from a primary ...
Esther Miriam Zimmer Lederberg (December 18, 1922 – November 11, 2006) was an American microbiologist and a pioneer of bacterial genetics.She discovered the bacterial virus lambda phage and the bacterial fertility factor F, devised the first implementation of replica plating, and furthered the understanding of the transfer of genes between bacteria by specialized transduction.
pBR322 is 4361 base pairs in length [1] and has two antibiotic resistance genes – the gene bla encoding the ampicillin resistance (Amp R) protein, and the gene tetA encoding the tetracycline resistance (Tet R) protein. It contains the origin of replication of pMB1, and the rop gene, which encodes a restrictor of plasmid copy number.
Replica plating is another technique used to plate out cells on agar plates. These four techniques are the most common, but others are also possible. It is crucial to work in a sterile manner to prevent contamination on the agar plates. [1] Plating is thus often done in a laminar flow cabinet or on the working bench next to a bunsen burner. [2]
An agar plate – an example of a bacterial growth medium*: Specifically, it is a streak plate; the orange lines and dots are formed by bacterial colonies.. A growth medium or culture medium is a solid, liquid, or semi-solid designed to support the growth of a population of microorganisms or cells via the process of cell proliferation [1] or small plants like the moss Physcomitrella patens. [2]
The results of Luria and Delbrück were confirmed in more graphical, but less quantitative, way by Newcombe. Newcombe incubated bacteria in a Petri dish for a few hours, then replica plated it onto two new Petri dishes treated with phage. The first plate was left unspread, and the second plate was then respread, that is, bacterial cells were ...
Replica plating is a technique that transfers colonies from one plate to another in the same spot as the last plate so the different media plates can be compared side by side. It is used to compare the growth of the same colonies on different plates of media to determine which environments the bacterial colony can or cannot grow in (this gives ...
In addition to β-galactosidase, pUC19 also encodes for an ampicillin resistance gene (amp R), via a β-lactamase enzyme that functions by degrading ampicillin and reducing its toxicity to the host. [5] Cells which have been successfully transformed with pUC19 can be differentiated from cells which have not by growing them on media with ampicillin.