Oct 14, 2020
Bacterial Culture for Plasmid Extraction
This protocol is a draft, published without a DOI.
- 1Chung Shan Medical University
Protocol Citation: iGEM-CSMU, Huan Jui Chang 2020. Bacterial Culture for Plasmid Extraction. protocols.io https://protocols.io/view/bacterial-culture-for-plasmid-extraction-bh45j8y6
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: July 01, 2020
Last Modified: October 14, 2020
Protocol Integer ID: 38781
Abstract
In this protocol, we present the method of culturing bacteria overnight after transformation. This protocol will typically undergo plasmid extraction and baterial storage in the ensuing steps.
Guidelines
The ratio between LB media and antibiotics varies under different scenarios. We showcase chloramphenicol in this protocol.
Safety warnings
Make sure that all the steps should be operated in laminar flow.
Before start
Follow the "preparation" section of protocol.
Preparation
Preparation
Turn on the UV light of the laminar flow for 00:10:00 to 00:15:00 in order to sterilize it.
Prepare LB Media.
Add 50 mL of LB Media to a 50 mL of centrifuge tube.
Add 50 μL of chloramphenicol.
Gently shake the tube to mix.
Prepare a LB Agar Plate with transformed bacterial colonies on it.
Protocol
Protocol
Add 7 mL of LB Media containing choramphenical to a 15 mL centrifuge tube
Use a toothpick to pick a single colony from a LB agar plate. Make sure the tip of the toothpick contains bacteria.
Drop the toothpick into the 15 mL centrifuge tube. Make sure the tip of toothpick fully submerge in the LB Media.
Incubate the 15 mL centrifuge tube for Overnight 16-18 hour .