Oct 17, 2022
Step A: Culturing
- k.z.cooper1
- 1University of Southampton
Protocol Citation: k.z.cooper 2022. Step A: Culturing. protocols.io https://dx.doi.org/10.17504/protocols.io.yxmvm2736g3p/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: In development
We are still developing and optimizing this protocol
Created: September 12, 2022
Last Modified: October 17, 2022
Protocol Integer ID: 69860
Abstract
Culturing step for bacterial genome resequencing project
Open the tracking form and verify you also have a copy of the batch tracking sheet printed out:
Preparation
Preparation
Ensure agar plates are pre-poured
Pre-label the plates using only the numbers provided on the spreadsheet under heading ‘Sample_Label’
5m
Get required glycerol stocks and keep on dry ice
20m
Creating streak plate
Creating streak plate
12h
12h
Ensure to cross reference with spreadsheet before every inoculation
Create streak plate inoculum using inoculation loop from glycerol stock
15m
Replace inoculation loop (after step I in the below figure) and dilute across the plate
Use 2 disposable loops, 1 for the initial step, then 1 for the remaining 4 steps
Incubate at 37°C overnight
12h
Creating LB broth overnight culture
Creating LB broth overnight culture
12h
12h
Pre-label tubes using numbers from spreadsheets
5m
Transfer 5 ml of LB broth into a universal tube
10m
Inoculate the LB broth with a single colony from the agar plate
15m
Grow at 37°C overnight at 180 rpm in a shaking incubator
12h
Place agar plates into fridge for storage