Apr 30, 2018
Version 2
Transformation of Vibrio natriegens V.2
- 1iGEM
Protocol Citation: Carlos Helbig 2018. Transformation of Vibrio natriegens. protocols.io https://dx.doi.org/10.17504/protocols.io.ptsdnne
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: April 27, 2018
Last Modified: April 30, 2018
Protocol Integer ID: 11858
Abstract
This protocol describes how to transform chemically competent Vibrio natriegens cells.
The protocol was described and published by Weinstock et al., 2016
Before start
You need:
- Chemically competent V. natriegens cells
- BHI + V2-Salts
10x V2-Salts:
| Chemical | Concentration | Molecular weight | Amount for 1L | |
| NaCl | 204mM | 58.44g/mol | 11.92176g | |
| KCl | 4.2mM | 74.55g/mol | 0.31311g | |
| MgCl2 | 23.14mM | 203.3g/mol | 4.704362g |
BHI:
For 1L medium 37g Brain heart infusion buillon
Defrost your competent cells on ice
Add your DNA template to the cells and mix gently
Incubate: 30min., on ice
Heatshock: 42°C, 45sec.
Incubate: 90sec., on ice
Add 1ml BHI + V2-Salts (pre-heated 30°C) to the cells
Incubate: 30°C, 1min.
Recover: 30°C, 120min., 200rpm
Centrifuge: 3000g, 4min.
Plate the pellet on warm agar plates (30°C)
Incubate: