Cloning Of A Cauliflower

1. Swab the laboratory bench with ethnol (70%)
2. Cut out a small piece of cauliflower- then roughly a centimetre across- with a sharp scalpel
3. Place in a petri dish, and using sharp cuts, divide the piece of cauliflower into three.
4. Carefully place the pieces of cauliflower, or explants, into 50cm of sodium hypochlorite (sodium (l) Chlorate), solution in a beaker.
5. Leave for 10 minutes
6. Undo the screw cap, and then flame the neck of the bottle of the sterile distilled water provided.
7. Pour out 50cm3 of sterile distilled water into three 100cm3 beaker
8. Flame the neck of the bottle and replace the cap
9. Dip the end of a pair of forceps into a beaker of ethanol, keeping this well away from the flame.
10.  Pass the forceps through a blue Bunsen flame to ignite the ethanol and sterilise them.
11.  Allow the forceps to cool, and then aseptically transfer the explants to a beaker of sterile water. Leave there for one minute.
12.  Rinse the explants twice more in the remaining two beakers of sterile distilled water. This rinsing process is critical to the growth of the explants. Leave in the beaker until required.
13. Using the hand opposite to the one you write with, remove the aluminium foil and cotton wool plug from the test tube of growth medium.
14. Flame the neck of the tube.
15. Using a pair of sterile forceps (flame as in stages 7 and 8), carefully, but quickly transfer one of the explants onto the surface of the growth medium in the tube.
16.  Carefully flame the neck of the tube and replace the cotton wool.
17. Cover the cotton wool with aluminium foil.
18. Repeat stages 15 to 17 with the other two explants, transferring them aseptically to two further tubes of growth media.
19. Keep the explants in a warm, light place.
20. Growth of the explants should be visible after about 10 days.
21. In this tube, aseptic technique has not been observed, and the explant has not grown.
22. Healthy explant and contaminated explant compared.

Safety Points

• Careful with scalpels
• Ethanol is flammable

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