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Week 12: August 22 to 27
August 22
WET LAB
Preparation of LB
We prepared additional 15 mL LB to continue activities from Aug 19, 2011
calculation:
15 mL de LB × (33μg Chloramphenicol)/(mL de LB) × mL/(34mg Chloramphenicol)= 14μL
We inoculate and incubate the petri dishes
August 23
WET LAB
We did a dirty miniprep to extract plasmidic DNA of the Biobricks
• BBa_K410000, pSB1C3
• BBa_K328003, pSB1C3
• BBa_K328001, pSB1C3
And stored until next day
August 24
WET LAB
Electrophoresis was applied to the following BioBricks:
• BBa_K410000, pSB1C3
• BBa_K328003, pSB1C3
• BBa_K328001, pSB1C3
1. Preparation of agarose 1%:
In a 250mL Erlenmeyer we mixed 0.5 g agarose with 50 mL of 1X TAE (for electrophoresis gel).
2. We heated in a microwave to dissolve (5 times of 20s each time stirring gently).
3. We cooled in a stream of gently flowing water.
4. We added 1,5 µL of ethidium bromide by touching the liquid. Stir until dissolved.
5. We added the blend to the electrophoresis cell with the combo on it. When the gel solidified we removed the comb and the wells were marked in the gel.
6. We placed part of the cell where the gel was solidified in the cell and filled it with TAE 1X.
7. We put on a piece of parafilm 10 drops of 2 μL c / u indicator LB 6X.
8. We placed in parafilm 2 µL of Supercoiled DNA Ladder reference.
9. We extracted 5 µL of an eppendorf and resuspended in LB 6X on parafilm.
10. Placed samples in the wells gell.
11. We repeated steps 9 and 10 until the contents of each eppendorf was placed in each well.
12. We resuspended 2 µL of supercoiled DNA ladder 2μL with a drop (2μL) LB 6X and we placed in the correspondant well.
13. We put the electrodes so that the samples ran from the negative to the positive terminal of the cell.
14. We ran the electrophoresis for about two hours.
Gel Electrophoresis of Part:BBa_K410000 and the CspA Promoter
1 2 3 4 5 6 7 8 9 10
1. Reference (DNA Supercoiled Ladder)
2, 3, 4 : BBa_K328001
5, 6, 7 : BBa_K328003
8, 9, 10: BBa_K410000
August 27
GENERAL MEETING SESSION
SAFETY DUE
This day we talk about all the safety information and documentation available in our country and institutions.
Final attribution of all the resposabilities of our team.
HUMAN PRACTICE
We were planning the content of the videos that we had decided to do in previous meetings, making the video really simple for the regular people and we prepared the contents about the iGEM history and the development of the SynBio.
We also prepared details of our project and ongoing activies.
Director of Session: Grimaldo E. Ureña & Lucia Palma.
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