Microbial Culturing
Microbial Culturing: As part of the Cloning and Sequencing Explorer Series.
Select for positive ligations and grow bacterial minipreps. Students use this module to prepare media for their microbiological needs for the Cloning and Sequencing Explorer Series.
Starter cultures are required for competent cell preparation. Students will make LB agar plates and streak them with E. coli bacteria for single colonies. Students then prepare LB broth to make an overnight starter culture from their streaked plate, which is used to prepare competent cells using the ligation and transformation module.
LB ampicillin agar plates are required to select for bacteria that have been transformed with plasmids — like pJET1.2 — which contain the Β-lactamase gene conferring ampicillin resistance to bacteria. After transformation, bacteria are plated onto LB ampicillin agar plates and incubated at 37°C and ampicillin resistant bacterial colonies will grow overnight.
LB broth containing ampicillin is required to culture minipreps grown from colonies transformed with ligated plasmid. Plasmids are then isolated from the minipreps using the Aurum plasmid mini purification kit.
