DNA fingerprinting made simple Cat no 130

1. DNA fingerprinting made simpleCat 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 A M 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 JeffreysInvented DNA fingerprinting

32. First ever DNA fingerprint

33. What it means m c f 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 CS S1 S2 S3 M

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