Mar 17, 2018
- 1Dalhousie University
Protocol Citation: Angela Tsai 2018. Heat Shock Transformation. protocols.io https://dx.doi.org/10.17504/protocols.io.nv5de86
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: March 17, 2018
Last Modified: March 28, 2018
Protocol Integer ID: 10909
Abstract
Purpose of this protocol is to introduce plasmid DNA into E. coli cells
Thaw desired E. coli strain on ice for ~10 minutes.
Add 5 uL of ligated DNA or ~10 ng of miniprepped plasmid DNA to 50 uL of E. colicells. Incubate on ice for 20-30 minutes.
Incubate bacteria for 45 seconds at 42°C.
After heat shock, let rest on ice for 10 minutes.
Add 250 uL LB Broth to E. coli solution.
Incubate, shaking, at 37°C for 1 hour.
After 1 hour, plate bacteria onto LB Agar+antibiotic plates, spreading solution evenly.
Incubate for 18-24 hours at 37°C.