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To create larger quantities of plasmids for isolation, liquid cultures are preferable to plates. Each colony on a plate in theory grew from a single bacterium. This means that all the cells in a single colony should be genetically identical. By putting at least one cell in a colony into liquid media, the cell should be able to grow and saturate the media overnight.
- Plates from transformation with individual colonies
- Liquid LB media in bacterial culture tube
- 1000X Kanamycin stock
- Add 1000X kanamycin to the LB so that the final concentration is 1X. Swirl to mix.
- Using a pipette tip, touch a single colony on the plate.
- Touch the pipette tip to the LB in the bacterial culture tube and lightly swirl. At least one cell must remain in the media.
- Grow at 37°C overnight while shaking.