Transformation Lab!

This lab was pretty neat! We went through a step-by-step process to figure out a different transformed version of bacteria. For instance we found out if they were more similar to the original non-transformed E. coli colonies. The genetically transformed bacterial cells were transformed after we did these steps (some pictures shown above):

1. Label one closed micro test tube +pGLO and another -pGLO
2. Open tubes and use transfer pipet to transfer 250 ul of CaC12
3. Place tubes on ice
4. Pick up colony of bacteria with sterile loop
5. Spin loop in +pGLO tube until colony is dispersed
6. Place back in ice
7. Repeat with -pGLO tube
8. Examine pGLO plasmid DNA solution with UV lamp
9. Immerse new sterile loop into the plasmid DNA stock tube and withdraw a loopful
10. Mix loopful into the cell suspension of +pGLO tube
11. Close and return to ice
12. Close -pGLO tube but DO NOT add plasmid DNA
13. Incubate tubes on ice for 10 minutes!
14. Put both tubes in water bath at 42 degrees C for 50 seconds
15. Place back in ice for 2 minutes
16. Remove and open a tube and use new sterile pipet to add 250 ul of LB nutrient broth
17. Reclose it and repeat with other tube
18. Incubate tubes for 10 minutes
19. Pipet 100 ul of the transformation and control suspension onto the plates
20. Spread suspensions evenly around surface
21. Stack plates and tape together
22. Stack and place upside down in 37 degrees C incubator until next day

When we were all done the results were pretty cool! It came out glowing all purple and what not! :) I was surprised because I didn't really think it would look all that cool, but I guess all those steps paid off! It looked awesome! :)