Team:UTP-Panama/Week 15

From 2011.igem.org

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===WET LAB===
===WET LAB===
-
(Objetives or Title):
+
Objetives:
-
--ESCRIBIR Y MEJORAR LAB--
+
'''DIRTY MINI PREP'''
 +
BBa-K381001 part
 +
Cultivation 9/6/2011
 +
1. Appropriately shook the tubes where the cultivations were.
 +
2. In each eppendorf 1.5mL of cultivation (9/6/2011) control (+)
 +
3. In each eppendorf 1.5mL of cultivation (9/6/2011) control (-)
 +
4. Centrifuged 13000rpm, 1 min
 +
5. Supernatant of eppendorf 6 tubes were eliminated
 +
6. Resuspended in 300µL buffer RNasa (P1)
 +
7. Added 300µL of buffer (P2)
 +
8. Mixed by inversion
 +
9. It was kept in room temperature during 5 minutes
 +
10. Added 300µL of buffer (P3)
 +
11. Mixed by inversion
 +
12. Kept in ice during 5 minutes
 +
13. Centrifuged 13000rpm, 10 min
 +
14. Extracted the supernatant and it was deposited in new eppendorf tubes carefully, avoiding to touch the pellets.
 +
15. Added to each tube 1000µL of PCA (Phenol:Chloroform:Isoamyl alcohol)
 +
16. Mixed by inversion
 +
17. Centrifuged 13000rpm, 5 min
 +
18. Transferred 750µL aqueous phase to new tubes properly labeled.
 +
19. Tubes with plasmids and isopropanol were centrifuged 13000rpm, 15 min
 +
20. Then isopropanol was removed
 +
21. Added 500µL EtOH 70% to wash it
 +
22. Mixed by inversion
 +
23. Again centrifuged 13000rpm, 5 min
 +
24. Eliminated EtOH 70% and were dried (air dry technique)
 +
25. Resuspended in 30µL of H20 ultra-pure (it was cold)
 +
26. Tubes were stored in a white box properly labeled
 +
 
 +
Note: Volumes of 500µL EtOH 70% and 30µL of H20 ultra-pure refer to that specific amount for each tube.
==September 14==
==September 14==

Revision as of 21:45, 28 September 2011


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Week 15: September 12 to 17

September 12

WET LAB

We grew the BBa-K381001 in liquid LB + chloramphenicol to then continue with DIRTY MINIPREP in order to extract its plasmids.

  1. Took out of the fridge 2 petri dishes with transformed bacteria with BBa-K381001
  2. It was used:
    1. 9.7µL of chloramphenicol
    2. and 10mL of liquid LB
  3. Prepared (+) and (-) controls for each petri dish in falcon tubes
  4. Each falcon tube of (+) control contains:
    1. 9.7µL of chloramphenicol/10mL of Liquid LB
    2. 5mL of liquid LB
    3. Aliquot of the cultivation
  5. The negative controls had almost the same content, excluding chloramphenicol
  6. The 4 falcon tubes were collocated in the incubator until next day


September 13

Scientific Comunication Activities

Presentation (by the students of iGEM UTP-Panama) in the Civil and Electrical Faculties about:
What's SynBio?
Applications of SynBio
What's iGEM?
What are we doing? (our project)
Future projects

WET LAB

Objetives: DIRTY MINI PREP BBa-K381001 part Cultivation 9/6/2011 1. Appropriately shook the tubes where the cultivations were. 2. In each eppendorf 1.5mL of cultivation (9/6/2011) control (+) 3. In each eppendorf 1.5mL of cultivation (9/6/2011) control (-) 4. Centrifuged 13000rpm, 1 min 5. Supernatant of eppendorf 6 tubes were eliminated 6. Resuspended in 300µL buffer RNasa (P1) 7. Added 300µL of buffer (P2) 8. Mixed by inversion 9. It was kept in room temperature during 5 minutes 10. Added 300µL of buffer (P3) 11. Mixed by inversion 12. Kept in ice during 5 minutes 13. Centrifuged 13000rpm, 10 min 14. Extracted the supernatant and it was deposited in new eppendorf tubes carefully, avoiding to touch the pellets. 15. Added to each tube 1000µL of PCA (Phenol:Chloroform:Isoamyl alcohol) 16. Mixed by inversion 17. Centrifuged 13000rpm, 5 min 18. Transferred 750µL aqueous phase to new tubes properly labeled. 19. Tubes with plasmids and isopropanol were centrifuged 13000rpm, 15 min 20. Then isopropanol was removed 21. Added 500µL EtOH 70% to wash it 22. Mixed by inversion 23. Again centrifuged 13000rpm, 5 min 24. Eliminated EtOH 70% and were dried (air dry technique) 25. Resuspended in 30µL of H20 ultra-pure (it was cold) 26. Tubes were stored in a white box properly labeled

Note: Volumes of 500µL EtOH 70% and 30µL of H20 ultra-pure refer to that specific amount for each tube.

September 14

WET LAB

(Objetives or Title): --ESCRIBIR Y MEJORAR LAB--

September 15

WET LAB

(Objetives or Title): --ESCRIBIR Y MEJORAR LAB--

September 16

WET LAB

(Objetives or Title): --ESCRIBIR Y MEJORAR LAB--

September 17

GENERAL SESSION

Morning: Talikng about WET LAB final activities. All the Team (under Human Practice Direction):

Schematization of algorithms and flow chart for a Human Practice Project.
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