Protocol Library

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E. coli Heat Shock Transformation

Developed by: opentrons

For this transformation, DNA is transferred into a tube of competent cells on the cold deck module via the p10 single channel pipette. After a 30 minute incubation, the competent cell mixture is transferred to a tube on the heat deck module via the p200 single channel pipette. After a 60 second pause, the competent cell mixture is transferred back to the cold deck for a 5 minute incubation. 200 uL of growth medium is then added to the mixture to complete the experiment.

You will need to manually complete any outgrowth incubation step(s) once the heat shock transformation is done (unless there is a second heating module on the deck). This protocol is flexible and can be easily adapted to suit each user’s needs. You can change the amount of DNA, growth media, number of samples and incubation times as necessary (see examples below).

Estimated time for this protocol is 40 minutes

Setup topTop Icon

  • CONTAINERS
    • A2 p200-rack
    • B1 p10-rack
    • B3 heat deck
    • C1 trash
    • D1 tube rack
    • D3 cold deck
  • PIPETTES
  • TIME ESTIMATION
    • ~40 minutes at 3,000 mm/sec
  • #TIPS USED
    • 1 x p10 tip
    • 2 x p200 tips

Click on each individual container to zoom in.

Source Code topTop Icon

Our protocols are written in a JSON file, please make sure that when you edit these files they are saved as a ".json".

Customize topTop Icon

If you’re using more than one sample, you’ll need to duplicate your transfer commands. This command moves the DNA sample from tube “B1”, to the competent cells in tube “A1” - make sure you change these well names when adding new transfer commands.

Line 131 shows a delay - this should be removed from all of your DNA transfer commands except the last one. Otherwise it will start a 30 minute delay after each DNA transfer, instead of adding all the DNA, and then beginning the cold incubation period.