Sep 10, 2021

Public workspaceGrowth of mixed E. coli colonies V.2

DOI
dx.doi.org/10.17504/protocols.io.bx3epqje
  • 1University of Exeter
Wolfram Moebius
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DOI: dx.doi.org/10.17504/protocols.io.bx3epqje
External link: https://doi.org/10.1128/mBio.01542-21
Protocol Citation: Wolfram Moebius 2021. Growth of mixed E. coli colonies . protocols.io https://dx.doi.org/10.17504/protocols.io.bx3epqjeVersion created by Wolfram Moebius
Manuscript citation:
Aranda-Díaz A, Rodrigues C, Grote A, Sun J, Schreck C, Hallatschek O, Souslov A, Möbius W, Huang KC, Bacterial Filamentation Drives Colony Chirality. mBio 12(6). doi: 10.1128/mBio.01542-21
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working.
Created: September 08, 2021
Last Modified: September 10, 2021
Protocol Integer ID: 53062
Abstract
Growth of mixed E. coli colonies on agar plates and between an agar plate and an agar pad
Make plates
Make plates
Autoclave Lennox LB (10 g/l tryptone / casein digest peptone, 5 g/l NaCl, 5 g/l yeast extract) with 1.5 % agar (w/v).
Optional: Add antibiotics at desired concentration after medium has cooled down sufficiently.
Pipette Amount10 mL of LB and agar in Petri dishes with diameter of Thikness6 cm .

Leave plates plates to dry DurationOvernight at TemperatureRoom temperature . Optionally enclose to avoid nonuniform drying.

Store and prepare plates
Store and prepare plates
If not used the day after pouring plates, store at Temperature4 °C . Enclose in plastic container or bag to avoid further drying of plates.

Before usage of plates that have been refrigerated, warm up plates at Temperature37 °C .

Prepare bacterial cultures
Prepare bacterial cultures
Grow overnight culture (colony picked or directly from glycerol frozen stocks) in Lennox LB.
Inoculate colonies and incubate
Inoculate colonies and incubate
Mix Amount900 µL Lennox LB and Amount50 µL of each of the two overnight cultures in an Eppendorf tube.
Vortex.

Inoculate Amount1 µL in centre of plate and let dry.
Optional: Cover colonies with agar pad. Cut about Thikness18 mm x Thikness18 mm agar pad inside plate. Use spatula to lift pad. Place vertically next to colony and let fall upside down onto colony. Use spatula to remove bubbles by pressing on top.

Place plates into container, together with sufficiently wet paper towels. Incubate for 7-8 days in a dark environment.