Oct 14, 2019

Public workspacetransformation of E.coli DH5α with plasmid DNA

DOI
dx.doi.org/10.17504/protocols.io.73fhqjn
  • 1Heinrich-Heine Universität Düsseldorf
Igem Dusseldorf
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DOI: dx.doi.org/10.17504/protocols.io.73fhqjn
Protocol Citation: Igem Dusseldorf 2019. transformation of E.coli DH5α with plasmid DNA. protocols.io https://dx.doi.org/10.17504/protocols.io.73fhqjn
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: October 09, 2019
Last Modified: October 14, 2019
Protocol Integer ID: 28487
Abstract
common transformation of E.coli DH5α with plasmid DNA
Materials
  • LB-Medium 1 ml (without antibiotics)
  • DH5a e.coli culture 50 µl
  • plasmid DNA 1 µl
  • plate with plasmid-specific antibiotics
  • Thermoblock (42°C)
  • ice bath
thaw competent cells on ice
add 1 µl plasmid DNA to cells
incubate cells on ice
30m
heat shock cells at 42°C
1m
cool on ice
2m
add 700 µl LB without antibiotics to the cells
incubate at 37°C (with shaking)
1h
plate 100 µl on antibiotic plates with Drigalski spatula
incubate over night at 37°C