Jun 28, 2022

Public workspaceE. coli Culture

DOI
https://dx.doi.org/10.17504/protocols.io.5jyl899o7v2w/v1
  • Bailey Clark1
  • 1Florida State University iGEM
Bec18
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DOI: https://dx.doi.org/10.17504/protocols.io.5jyl899o7v2w/v1
Protocol Citation: Bailey Clark 2022. E. coli Culture. protocols.io https://dx.doi.org/10.17504/protocols.io.5jyl899o7v2w/v1
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: February 11, 2022
Last Modified: June 09, 2024
Protocol Integer ID: 58052
Keywords: abstract, culture
Abstract
The abstract will be added later.
Materials
  • Amount3 mL LB medium
  • Appropriate Antiobiotic
  • 15 mL tube
  • Thermomixer
  • Agar plate with bacteria colonies
  • 1 µL inocculation loop





Troubleshooting
Before start
A video if unfamiliar: https://youtu.be/TWt62pgAEh0
Medium and Antibiotic Preparation
5m
Add Amount2 mL (Amount3 mL also acceptable) of LB medium and the appropriate antibiotic that selects for the target strain into a 15 mL tube.
Protocol
NAME

Kanamycin 50 mg/mL Stock Solution 

CREATED BY
Bec18


Place the 15mL tube in the ThermoMixer at 37°C, 300 rpm, for Duration00:05:00 .
Equipment
ThermoMixer® C
NAME
Eppendorf
BRAND
Catalog No. 2231000680
SKU
https://online-shop.eppendorf.us/US-en/Temperature-Control-and-Mixing-44518/Instruments-44519/Eppendorf-ThermoMixerC-PF-19703.html
LINK


5m
Retrieving Cells
Gather cells.
Swab a single colony on the agar plate with a sterile 1µL inoculation loop.

OR Dip a sterile 10 µL innoculation loop in a preexisting culture tube.

Innnoculation
Innoculate the 15 mL tube with the loop.
Place the 15 mL tube in the thermomixer for 16 hours at 37°C and 300 rpm.