1. DNA fingerprinting made simple Cat no 130 Edvotek Europe

2. What are we doing today? Introduction Prepare gels for loading Load samples Run gels Stain & analyse results

3. Why study DNA?

5. Ready to load kit 130 DNA fingerprinting made simple –DNA based electrophoreis –Needs staining to see result

6. Health & safety No toxic chemicals Safe solutions and reagents used in place of research materials

7. Compare samples Use electrophoresis Micropipette to load samples Electrical current will separate by size Pattern is DNA fingerprint

8. Gel casting

9. Running buffer TAE buffer 20ml 50x buffer to 1 litre with water Distilled water ideal but tap ok Can be reused a few times Store unused for future use ok

10. Agarose 0.8% Agarose 3g in 375 ml dilute buffer Melt in microwave/autoclave Pour when hand hot

11. Agarose Store gels for 1-2 weeks in fridge wrapped in cling film or plastic bag Keep any left over gels and remelt next time

12. The experiment

13. Remove comb & ends

15. Fixed volume minipipette

16. Adjustable micropipette

17. Dry loading You can load gel dry instead of through buffer! Otherwise load “wet” through buffer Either way, remember Do not puncture bottom of well Change tips between each sample

19. HexaGel and EVT 300 power supply

20. 130 kit gel Loading Load 40  l of each sample into the wells AMAM B CS C S1 D S2 E S3 F

21. Run gels Put on lid and attach to power supply Run for 30 minutes at 150 volts Or 75 volts for 40-50 minutes Check for bubbles at electrode Run until tracking dye halfway across gel

22. Background on DNA fingerprinting

23. How do you make a DNA fingerprint Get DNA from someone Cut it up with enzymes Separate different size pieces on a gel Pattern is a DNA fingerprint

24. Get some DNA

25. DNA is very long 2 metres per cell! 3 billion letters long (ACTG) Like a piece of string

26. Cut with scissors

27. Where to cut? Decided by combination of letters GTTAAC is cut by one sort of enzyme TTAA cut by a different sort

28. Real enzymes

29. Separate on gel Made of seaweed - agarose Different size pieces move through mesh - like a sieve Small move quicker than big Called DNA electrophoresis!

30. move by size big medium small

31. Prof. Sir Alec Jeffreys Invented DNA fingerprinting

32. First ever DNA fingerprint

33. What it means mcf M = mother C = child F = father Child has bands in Common with mother & father

34. After gel run stain the gel

35. Stain 130 kit gel MetBlue card blue side down 5 mins Weigh with gel tray Destain in warm water 10-15 mins Leave overnight in fridge for best result Keep long term in bag in fridge

36. Stain Gel

38. Gel storage Can store gels in fridge before staining over weeknight or weekend Cannot store dye kits overnight before viewing as diffuse!

39. 130 kit result M CSS1S2S3

40. Thank you DNA experiments can be carried out easily Fun and relevant to wider world Promote understanding