Copyright © Dean Madden, 2013

The bacterium Bacterial chromosome Plasmids Cytoplasm Ribosomes Cell wall Cell membranes

The plasmid Origin of replication this is needed so that the plasmid can be copied, using the bacterium’s DNA copying machinery Kanamycin resistance gene the enzyme encoded by this gene stops the antibiotic kanamycin from working GFP gene This gene encodes green fluorescent protein, which glows in UV light

Aequorea victoria

GFP

Glo Fish

These should be grown 1–2 days in advance Stock bacteria

Twiddle vigorously to dislodge the bacteria and disperse them Transformation buffer

Leave on ice for 15 minutes to recover

Bacterial cell with pores in cell membrane GFP plasmid DNA Bacterial cell with pores in cell membrane

3 drops (100 µL) LB agar + kanamycin + IPTG

Seal the plates with tape Label the base of each plate with your initials, the date and the type of bacteria used Seal the plates with tape

TRANSFORMED CELLS MUST BE DESTROYED AFTER USE BIOHAZARD TRANSFORMED CELLS MUST BE DESTROYED AFTER USE