Corum:Plating
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Overview
Plating of E. coli cells onto an LB-agar plate.
Materials
- LB-agar plate (w/ appropriate antibiotic, if required)
- sterilized, wooden stick or sterile Pasteur pipette
- E. coli cell sample, either from another plate, frozen stock, or cell culture
Procedure
- From plate or frozen stock:
- Under sterile conditions, pick a single colony using a sterile stick (if frozen stock, dip the tip of the stick into the stock) and drag the cells along the surface of the plate.
- Using another stick, drag through the line of (now-deposited cells) to an untouched portion of the plate surface.
- Repeat the previous step on the newly formed line until there are 4-5 total lines on the plate.
- Incubate overnight 37 °C (or special temperature).
- From cell culture:
- Dilute the cell sample to a density of ~104 cells / mL (OD600 ~ 10-5).
- Under sterile conditions, pipette 30 μL cells onto the LB-agar plate. Spread until dry using the sterile Pasteur pipette. (This will provide ~300 colonies.)
- Incubate overnight 37 °C (or special temperature).
Notes
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Contact
Digital Signature
- SC 17:27, 25 June 2012 (EDT):
or instead, discuss this protocol.