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Bacterial
Transformation
National Centre for Biotechnology Education
Copyright © Dean Madden, 2013
www.ncbe.reading.ac.uk
The bacterium
Bacterial
chromosome
Plasmids
Cytoplasm
Ribosomes
Cell wall
Copyright © Dean Madden, 2013
Cell
membranes
The plasmid
Kanamycin
resistance
gene
the enzyme
encoded by
this gene
stops the
antibiotic
kanamycin
from working
Origin of
replication
this is needed so
that the plasmid
can be copied,
using the
bacterium’s DNA
copying machinery
Copyright © Dean Madden, 2013
GFP gene
This gene encodes green
fluorescent protein, which
glows in UV light
Copyright © Dean Madden, 2013
Aequorea victoria
GFP
Copyright © Dean Madden, 2013
Copyright © Dean Madden, 2013
Glo Fish
Copyright © Dean Madden, 2013
Copyright © Dean Madden, 2013
Copyright © Dean Madden, 2013
These should be grown
1–2 days in advance
Copyright © Dean Madden, 2013
Stock bacteria
Twiddle vigorously
to dislodge the
bacteria and
disperse them
Copyright © Dean Madden, 2013
Transformation
buffer
Leave on ice for 15
minutes to recover
Copyright © Dean Madden, 2013
GFP
plasmid
DNA
Copyright © Dean Madden, 2013
Bacterial cell with pores
in cell membrane
3 drops
(100 µL)
Copyright © Dean Madden, 2013
LB agar
+ kanamycin
+ IPTG
Copyright © Dean Madden, 2013
Label the base of each
plate with your initials, the
date and the type of
bacteria used
Seal the
plates with
tape
Copyright © Dean Madden, 2013
Copyright © Dean Madden, 2013
Copyright © Dean Madden, 2013
Copyright © Dean Madden, 2013
Copyright © Dean Madden, 2013
Copyright © Dean Madden, 2013
BIOHAZARD
TRANSFORMED CELLS
MUST BE DESTROYED
AFTER USE
Copyright © Dean Madden, 2013
Before
Copyright © Dean Madden, 2013
After
Copyright © Dean Madden, 2013
Copyright © Dean Madden, 2013