Team:Cambridge/Protocols/TransformationofB.subtilis

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Transformation of Bacillus subtilis

A technique used to introduce foreign DNA into competent Bacillus cells.

Theory

Bacillus subtilis is a rod-shaped, soil-dwelling bacterium . It is widely used as a model organism for studying biology of Gram-positive bacteria which, in contrast to Gram-negative E.coli, do not possess the outer membrane over the peptidoglycan cell wall. Additionally, Bacillus is naturally competent, which means that no sophisticated pre-treatment is needed for transformation with foreign DNA.

Practice

  • Add to a clean Eppendorf tube:
Reagent Quantity
competent Bacillus cells 400μl
DNA 3μl
  • Incubate at 37ºC for 1 hour.
  • Plate on an agar plate containing antibiotic, which allows for selection of successful transformants.

Safety

All equipment (including gloves) that may have come into contact with the bacteria must be autoclaved for decontamination purposes.



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