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National Centre for Biotechnology Education www.ncbe.reading.ac.uk Bacterial Transformation Copyright © Dean Madden, 2013.

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Presentation on theme: "National Centre for Biotechnology Education www.ncbe.reading.ac.uk Bacterial Transformation Copyright © Dean Madden, 2013."— Presentation transcript:

1 National Centre for Biotechnology Education Bacterial Transformation Copyright © Dean Madden, 2013

2 The bacterium Bacterial chromosome Plasmids Ribosomes Cell wall Cell membranes Cytoplasm

3 Copyright © Dean Madden, 2013 GFP gene This gene encodes green fluorescent protein, which glows in UV light 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 bacteriums DNA copying machinery The plasmid

4 Copyright © Dean Madden, 2013 Aequorea victoria

5 Copyright © Dean Madden, 2013 GFP

6 Copyright © Dean Madden, 2013 Glo Fish

7 Copyright © Dean Madden, 2013

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

11 Copyright © Dean Madden, 2013 Twiddle vigorously to dislodge the bacteria and disperse them Transformation buffer

12 Copyright © Dean Madden, 2013 Leave on ice for 15 minutes to recover

13 Copyright © Dean Madden, 2013 Bacterial cell with pores in cell membrane GFP plasmid DNA

14 Copyright © Dean Madden, 2013 LB agar + kanamycin + IPTG 3 drops (100 µL)

15 Copyright © Dean Madden, 2013

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

17 Copyright © Dean Madden, 2013

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22 BIOHAZARD TRANSFORMED CELLS MUST BE DESTROYED AFTER USE

23 Copyright © Dean Madden, 2013 Before

24 Copyright © Dean Madden, 2013 After

25 Copyright © Dean Madden, 2013


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