Sep 23, 2019
Chemically Competent Cells
- 1Wageningen University
- iGEM Wageningen 2019
Protocol Citation: Alba Balletbó 2019. Chemically Competent Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.7jwhkpe
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: September 23, 2019
Last Modified: September 23, 2019
Protocol Integer ID: 27990
Materials
MATERIALS
CaCl2
LB mediumCatalog #/
Culture desired strain O/N 0.5 mL/50 mL LB in a Greiner tube.
Grow to OD600 0.6 in 0.1/10 mL LB.
Centrifuge at 8000rpm for 5 minutes.
Resuspend in 5mL ice-cold CaCl2.
Distribute 1.5 mL over 3 Eppendorfs and spin for 30 seconds, 8000rpm.
Resuspend pellet in 0.5 mL ice-cold CaCl2, flick the tube gently.
Aliquot into 50µL Cryo-tubes and store at -80ºC until transformation.