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Team:Colombia/Notebook
From 2011.igem.org
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Revision as of 15:47, 26 September 2011
Template:Https://2011.igem.org/User:Tabima
Contents |
Colombia @ iGem Notebook
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).
July 19
- Pass strain of Vibrio fischeri to blood agar base.
- Check the growth of the isolates
- Pass the transformed bricks 6, 7, 9 and 10.
- Pass the isolates the solid media to liquid media (2,4,5 and 8)
July 21
- Digestion to confirm No. 1, 3 and 4
| Reactives | 1X | 6X |
| H2O | 29,4µL | 160,4 µL |
| Buffer N. 3 | 4 µL | 24 µL |
| EcoRI | 0,3 µL | 1,8 µL |
| PstI | 0,3 µL | 1,8 µL |
| DNA | 7 µL | 40 µL |
July 22
- Minipreps
- Electrophoresis of the digestions
- Task:
- Confirm minipreps
- Re-suspend primers
July 27
- To prepare LB and SOC medium and autoclaved
- The bricks 7, 9 and 10 were again transformed and plated
- Plate the brick 6.
July 30
- Minipreps with RNase.
- Agarose gel Electrophoresis—Results were not obtained. REPEAT!!
August
August 3
- PCR 16S to DNA Vibrio fischeri U. Nacional
- To expected a band of 1400 bp.
| 1X | 3X | |
| H2O | 6,8µL | 20,4µL |
| Bµffer | 1µL | 3µL |
| MgCl2 | 0,8µL | 2,4µL |
| dNTPs | 0,2µL | 0,6µL |
| Fw7 | 0,2µL | 0,6µL |
| Rv49 | 0,2µL | 0,6µL |
| Taq | 0,1µL | 0,3µL |
| DNA | 1µL | - |
| 10µL |
