From 2011.igem.org
Daily iGemWork
Here you can find our daily work in the Lab!
June
June 30:
- Biobricks 1, 2, 3, 4 and 5 were resuspended
- Miniprep Solutions (I, II and III) were prepared.
July
July 1
- Biobricks 1, 2, 3, 4 and 5 were resuspended.
July 5
- Biobrick 2 presented no colonies.
- Colonies from bricks 1, 3, 4 and 5 were stinged.
Task:
To make LB medium (15x25mL)
July 6
- Biobricks 1, 3 and 4 were twice plated.
- Brick 5 didn't grow up.
- We've added Ampiciline (3.75mL) and Kanamicine (1.875mL) to the boxed from the previous day.
- LB medium was prepared (15x25mL).
- Liquid LB was prepared (400mL).
Task:
To add ampiciline and tetracicline to the boxes.
Claim the liquid LB
Scrape ....
Sting biobricks 2 and 5 (no clons)
Pick up the tubes with Merceditas
July 7
- Clons 2 and 5 didn't work out.
- Clons 1, 3 and 4 were planted in LB liquid.
- E. Coli inoculation in Coffee
- Kanamicine resistence plasmid: 0.2 optic density.
- Direct inoculation x 2 and C(-) MgCl2.
Task:
Print electroporation protocol.
Ask Juan D. Olarte about the inoculation in coffee.
Minipreps for confirmation of 1, 3 and 4.
Competent cells for clons 2 and 5.
July 8
- Minipreps for 1, 3 and 4 were made.
- Plant sheets of coffee into different plates: Each sheet was cut in the middle and they were incubated at 25°C and 37°C LB+Kan.
Task:
To finish the minipreps from the addition of RNAsa.
Check the E. Coli growth in coffee.
July 11
- Minipreps have been finished. They've been planted in gel: Low concentration (25 ng/uL).
- Transformation protocol in chemical cells.
Task:
Prepare 250 mL of SOC medium
Print the Transformation protocol for chemical cells.
July 12
- Strains 1 and 4 have been conserved.
- LB liquid culture was prepared again for brick 3.
- E. Coli growth results.
Task:
Print the Transformation protocol for chemical cells.
Competent cells for clons 2 and 5.
Preserve brick 3.
Confirm bricks 1, 3 and 4 (Digestion)
Resuspend all Biobricks.
Check the E. Coli growth in coffee.
July 14
- E. Coli didn't grow up on the sheets.
- Chemical competent cells were made (DH5α).
- Brick 3 was left to grow in liquid medium.
- Bricks 2, 5, 6,7, 8, 9 and 10 were transformed and plated.
Task:
Print the Transformation protocol for chemical cells.
Preserve brick 3.
Confirm all biobricks (Digestion)
Sting the transformed bricks.
Add antibiotic to the mediums made today.
July 15
- Brick 3 was preserved in Revco.
- Bricks 2, 5 and 8 were stinged.
- 25 LB+Kan boxes x 25 mL.
- No colonies in brick 6.
- Bricks 7, 9 and 10 were contaminated.
Task:
Print the chemical cells protocol.
Confirm all biobricks (Digestion).
August