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Nesta DIYbio

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Nesta DIYbio

  1. 1. Biohacking! (AKA Synthetic Biology for Beginners)Sunday, 10 February 13
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  9. 9. Lab Rules • No food & drink in the lab • Gloves & aprons at all times • Wash/gel your hands when you leave the lab • Aargh spills! • No running, diving or bombingSunday, 10 February 13
  10. 10. Aseptic Technique • Keep air exposure to a minimum. • Don’t cross-contaminate.Sunday, 10 February 13
  11. 11. Micropipetting • Two different types • Set the volume by turning the dial • Suck! • Squirt! • Remember: Always use a fresh tip.Sunday, 10 February 13
  12. 12. …Sunday, 10 February 13
  13. 13. 1. • Iron Microbeads! (In tube 1) • Wash them… 50µl of washing buffer. Mix. • Clean them… pull the beads to one side with the magnet; extract the liquid. • Wash and clean them again…Sunday, 10 February 13
  14. 14. 2. • Ligation! Glues the first “part” to the beads. • Add 7µl of ligation mix (tube with the spot on) and mix. • Transfer this to the tube containing our first part (Tube 2) • Wait 5-10 minutes…Sunday, 10 February 13
  15. 15. 3. • Wash them… 50µl of washing buffer. Mix. • Clean them… pull them to one side with the magnet; extract the liquid. • Wash and clean them again…Sunday, 10 February 13
  16. 16. 4. • Ligation again! Glue the final two parts to our beads. • Add 7µl of ligation mix (tube with the spot on) and mix. • Transfer this to the tube containing our second part (Tube 3) and mix. • Transfer this to the tube containing our first part (Tube 4) and mix some more. • Wait 5-10 minutes…Sunday, 10 February 13
  17. 17. 5. • Wash them… 50µl of washing buffer. Mix. • Clean them… pull them to one side with the magnet; extract the liquid. • Wash and clean them again…Sunday, 10 February 13
  18. 18. 5. • Elution! Gets rid of the beads • Add 20µl of elution buffer and mix • Pull the beads to one side with the magnet • Extract the finished part in liquid form; put it into one of the empty tubes.Sunday, 10 February 13
  19. 19. …Sunday, 10 February 13
  20. 20. Modify that e.coli! • Put 100µl of Transformation buffer in an empty tube; put the tube on ice for 5 minutes, then… • Take a loop. Grab some e.coli and add it to the transformation buffer – “twiddle vigorously”. Wait for 10 minutes… • Add 40µl of your creation and mix. • Bring your tube to the front!Sunday, 10 February 13
  21. 21. Modify that e.coli! (Pt. 2) • “Heat shock” our e.coli for exactly 30 seconds in the water bath (42ºc) • Put them back on ice for 1-2 minutes • Add 100µl of recovery broth, keep them warm in your hand until…Sunday, 10 February 13
  22. 22. Modify that e.coli! (Pt. 3) • Put 100µl of your modified e.coli in a petri dish. • Remove the spreader from the alcohol and flame it (so there’s no alcohol left) • Spread the e.coli around your plate • …and that’s it! Bring your plate to the front and put it in the incubator.Sunday, 10 February 13

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