Team:Newcastle/Transformation of E. coli


iGEM Homepage Newcastle University BacillaFilla Homepage Image Map

Transformation of E. coli

Materials required

  • E. coli DH5α (200 µl)
  • Appropriate vector DNA
  • Heat block
  • Bucket of ice
  • Pipettes
  • Eppendorf tubes
  • 1.5% agar plate containing appropriate antibiotics


  1. Thaw a 200 µl aliquot of E. coli DH4α and add the transforming DNA (10 ng of vector DNA in 10 µl).
  2. Incubate on ice for 30 minutes.
  3. Heat-shock the cells for 120 seconds at 42°C, and place on ice again for 3-4 minutes.
  4. Add 1 ml of LB broth and incubate the cells at 37°C for 1-1.5 hr in a water bath with gentle shaking.
  5. Plate 200 µl of transformed E. coli onto 1.5% agar plate containing the appropriate selection markers.
  6. Incubate the plates overnight at 37°C.

Go back to our Protocol List

Newcastle University logo.png    Newcastle cbcb logo.pngNewcastle Biomedicine logo.gif    Team Newcastle CEG logo.gif
Newcastle iww logo.jpg  UNIPV Pavia Logo.gif  Newcastle BBSRC.gif    Newcastle Genevision logo.png Newcastle WelcomeTrust.jpg
FaceBook Icon